Anti-tumor compound and preparation method and use thereof

ABSTRACT

The present application relates to an anti-tumor compound and a preparation method and use thereof, and in particular to a compound or a tautomer, a mesomer, a racemate, an enantiomer or a diastereoisomer thereof, or a mixture thereof, or a pharmaceutically acceptable salt thereof, and a preparation method and use thereof.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Continuation of and claims priority under 35U.S.C. § 111 to Patent Cooperation Treaty application PCT/CN2021/121721,filed Sep. 29, 2021, which claims the benefit of Chinese PatentApplication No. 202011061580.7, filed Sep. 30, 2020, priority is claimedto both of these applications and the disclosures of these priorapplications are considered part of the disclosure of this applicationand to the extent allowed the entire contents of the aforementionedapplications are incorporated herein.

SEQUENCE LISTING

-   -   This application incorporates by reference in its entirety the        Sequence Listing entitled        “2022-10-07_262790-511702_ST25amended.txt” is 36,936 bytes in        size and was created on Oct. 10, 2022, and filed electronically        herewith.

TECHNICAL FIELD

The present application relates to the field of biomedicine, and inparticular to an anti-tumor compound and a preparation method and usethereof.

BACKGROUND OF THE INVENTION

Currently, small molecules with cytotoxicity for antibody-drugconjugates (ADCs) can be camptothecin derivatives, which produce ananti-tumor effect by inhibiting topoisomerase I. Camptothecinderivatives can be used in ADCs. There is still a need to developcamptothecin derivatives and ADC drugs with better therapeutic effectand/or safety.

SUMMARY OF THE INVENTION

The present application provides a compound or a tautomer, a mesomer, aracemate, an enantiomer or a diastereoisomer thereof, or a mixturethereof, or a pharmaceutically acceptable salt thereof, which may haveone or more effects selected from the group consisting of: (1) havinginhibitory activity against in vitro proliferation of tumor cells; (2)having targeting inhibition; (3) having plasma stability; (4) having invivo tumor inhibiting effect; (5) having bystander effect; (6) havingcapacity in inhibiting transport via a transporter; (7) having in vivotumor targeting capability; and (8) having good in vivo safety.

In one aspect, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound comprises a structure shown as formula(II-A):

wherein, X¹ is saturated C, and X¹ is substituted with R^(n);

ring A is selected from the group consisting of: 3-10 membered saturatedor partially unsaturated heterocyclyl and 3-10 membered saturated orpartially unsaturated carbocyclyl, wherein ring A is substituted with 0or at least 1 substituent R^(1a);

when ring A is 3-10 membered saturated or partially unsaturatedcarbocyclyl, ring A is substituted with p L², and L² is not R^(n);

or, when ring A is 3-10 membered saturated or partially unsaturatedheterocyclyl, ring A is substituted with p L²;

L² is —R²-L³-, and R² is used for direct or indirect linking of aligand;

L³ is —(C(R^(3a))(R^(3b)))_(m)—, wherein when L³ comprises a methyleneunit, 0 or at least 1 methylene unit of L³ is independently replaced by—N(R⁴)C(O)—, —C(O)N(R⁴)—, —C(O)—, —OC(O)—, —C(O)O—, —NR⁴—, —O—, —S—,—SO—, —SO₂—, —P(R⁴)—, —P(═O)(R⁴)—, —N(R⁴)SO₂—, —SO₂N(R⁴)—, —C(═S)—,—C(═NR⁴)—, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

R² is selected from the group consisting of: —O—, —(R^(2a))N—, —S— and—P(═O)(R^(2a))—;

L¹ is —(C(R^(5a))(R^(5b)))_(n)—, wherein when L¹ comprises a methyleneunit, 0 or at least 1 methylene unit of L¹ is independently replaced by—N(R⁶)C(O)—, —C(O)N(R⁶)—, —C(O)—, —OC(O)—, —C(O)O—, —NR⁶—, —O—, —S—,—SO—, —SO₂—, —P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—, —SO₂N(R⁶)—, —C(═S)—,—C(═NR⁶)—, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

wherein each R^(1a), each R^(2a), each R^(3a), each R^(a3), each R⁴,each R^(5a), each R^(5b), each R⁶ and each R^(n) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R,or a C₁₋₆ aliphatic group optionally substituted with R;

wherein each R, each R^(a) and each R^(b) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

m and n are each independently selected from the group consisting ofintegers ≥0, and p is an integer ≥1.

In one aspect, the present application provides a compound of generalformula (II-E_(x)) or a tautomer, a mesomer, a racemate, an enantiomeror a diastereoisomer thereof, or a mixture thereof, or apharmaceutically acceptable salt thereof,

wherein, X¹ is saturated C, and X¹ is substituted with R^(n);

ring A is selected from the group consisting of: 3-10 membered saturatedor partially unsaturated heterocyclyl and 3-10 membered saturated orpartially unsaturated carbocyclyl, wherein ring A is substituted with 0or at least 1 substituent R^(1a);

when ring A is 3-10 membered saturated or partially unsaturatedcarbocyclyl, ring A is substituted with p L², and L² is not R^(a);

or, when ring A is 3-10 membered saturated or partially unsaturatedheterocyclyl, ring A is substituted with p L²;

L² is —R²-L³-, and R² is used for direct or indirect linking of aligand;

L³ is —(C(R^(3a))(R^(3b)))_(m)—, wherein when L³ comprises a methyleneunit, 0 or at least 1 methylene unit of L³ is independently replaced by—N(R⁴)C(O)—, —C(O)N(R⁴)—, —C(O)—, —OC(O)—, —C(O)O—, —NR⁴—, —O—, —S—,—SO—, —SO₂—, —P(R⁴)—, —P(═O)(R⁴)—, —N(R⁴)SO₂—, —SO₂N(R⁴)—, —C(═S)—,—C(═NR⁴)—, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

R² is selected from the group consisting of: —O—, —(R^(2′))N—, —S— and—P(═O)(R^(2a))—;

L¹ is —(C(R^(5a))(R^(5b)))^(n)—, wherein when L¹ comprises a methyleneunit, 0 or at least 1 methylene unit of L¹ is independently replaced by—N(R⁶)C(O)—, —C(O)N(R⁶)—, —C(O)—, —OC(O)—, —C(O)O—, —NR⁶—, —O—, —S—,—SO—, —SO₂—, —P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—, —SO₂N(R⁶)—, —C(═S)—,—C(═NR⁶)—, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

wherein each R^(1a), each R^(2a), each R^(3a), each R^(3b), each R⁴,each R^(5a), each R^(5b), each R⁶ and each R^(n) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R,or a C₁₋₆ aliphatic group optionally substituted with R;

wherein each R, each R^(a) and each R^(b) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

m and n are each independently selected from the group consisting ofintegers ≥0, and p is an integer ≥1.

In one aspect, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound comprises a structure shown as formula(II-C_(x)):

wherein, L is -L_(a)-L_(b)-L_(c)-;

-L_(a)- is selected from the group consisting of:

wherein W is —(C(R^(wa))(R^(wb)))_(wn)—, Y is —(OCH₂CH₂)_(yn)—O_(yp)—,and Z is —(C(R^(za))(R^(zb)))_(zn);

wherein wn is selected from the group consisting of integers ≥0, and

0 or at least 1 methylene unit of W is independently replaced by -Cyr-,—N(R^(wx))C(O)—, —C(O)N(R^(wx))—, —C(O)—, —OC(O)—, —C(O)O—, —NR^(wx)—,—O—, —S—, —SO—, —SO₂—, —P(R^(wx))—, —P(═O)(R^(wx))—, —N(R^(wx))SO₂—,—SO₂N(R^(wx))—, —C(═S)—, —C(═NR^(wx))—, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

wherein yn is selected from the group consisting of integers ≥0, and ypis 0 or 1;

wherein zn is selected from the group consisting of integers ≥0, and

0 or at least 1 methylene unit of Z is independently replaced by -Cyr-,—N(R^(zx))C(O)—, —C(O)N(R^(zx))—, —C(O)—, —OC(O)—, —C(O)O—, —NR^(zx)—,—O—, —S—, —SO—, —SO₂—, —P(R^(zx))—, —P(═O)(R^(zx))—, —N(R^(zx))SO₂—,—SO₂N(R^(zx))—, (═S)—, —C(═NR^(zx))—, —N═N—, —N═C— or —C(═N₂)—;

-Cyr- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene and 3-10membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or independently substituted with at least 1substituent R^(cx);

wherein each R^(wa), each R^(wb), each R^(za), each R^(zb), each R^(wx),each R^(zx) and each R^(cx) are each independently hydrogen, protium,deuterium, tritium, halogen, —NO₂, —CN, —OR^(r), —SR^(r),—N(R^(ra))(R^(Rrb)), —C(O)R^(r), —CO₂R^(r), —C(O)C(O)R^(r),—C(O)CH₂C(O)R^(r), —S(O)R^(r), —S(O)₂R^(r), —C(O)N(R^(ra))(R^(rb)),—SO₂N(R^(ra))(R^(rb)), —OC(O)R^(r), —N(R)SO₂R^(r), or a C₁₋₆ aliphaticgroup optionally substituted with R^(r);

wherein each R^(r), each R^(ra) and each R^(rb) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

-L_(b)- represents a peptide residue consisting of 2 to 7 amino acids;

-L_(c)- is selected from the group consisting of:

wherein R^(L1) and R^(L2) are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO₂,—CN, —OH, —SH, —NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H,—S(O)₂H, —C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H and a C₁₋₆ aliphaticgroup;

wherein, X¹ is saturated C, and X¹ is substituted with IV;

ring A is selected from the group consisting of: 3-10 membered saturatedor partially unsaturated heterocyclyl and 3-10 membered saturated orpartially unsaturated carbocyclyl, wherein ring A is substituted with 0or at least 1 substituent R^(1a);

when ring A is 3-10 membered saturated or partially unsaturatedcarbocyclyl, ring A is substituted with p L², and L² is not IV;

or, when ring A is 3-10 membered saturated or partially unsaturatedheterocyclyl, ring A is substituted with p L²;

L² is —R²-L³-, and R² is used for direct or indirect linking of aligand;

L³ is —(C(R^(3a))(R^(3b)))_(m)—, wherein when L³ comprises a methyleneunit, 0 or at least 1 methylene unit of L³ is independently replaced by—N(R⁴)C(O)—, —C(O)N(R⁴)—, —C(O)—, —OC(O)—, —C(O)O—, —NR⁴—, —O—, —S—,—SO—, —SO₂—, —P(R⁴)—, —P(═O)(R⁴)—, —N(R⁴)SO₂—, —SO₂N(R⁴)—, —C(═S)—,—C(═NR⁴)—, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

R² is selected from the group consisting of: —O—, —(R^(2a))N—, —S— and—P(═O)(R^(2a))—;

L¹ is —(C(R^(5a))(R^(5b)))_(n)—, wherein when L¹ comprises a methyleneunit, 0 or at least 1 methylene unit of L¹ is independently replaced by—N(R⁶)C(O)—, —C(O)N(R⁶)—, —C(O)—, —OC(O)—, —C(O)O—, —NR⁶—, —O—, —S—,—SO—, —SO₂—, —P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—, —SO₂N(R⁶)—, —C(═S)—,—C(═NR⁶)—, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

wherein each R^(1a), each R^(2a), each R^(1a), each R^(ab), each R⁴,each R^(5a), each R^(5b), each R⁶ and each R^(n) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R,or a C₁₋₆ aliphatic group optionally substituted with R;

wherein each R, each R^(a) and each R^(b) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

m and n are each independently selected from the group consisting ofintegers ≥0, and p is an integer ≥1.

In one aspect, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound comprises a structure shown as formula(II-Dx):

wherein, Ab is a ligand, and an average connection number N^(a) is aninteger or a decimal from 1 to 10;

L is -L_(a)-L_(b)-L_(c)-;

-L_(a)- is selected from the group consisting of:

wherein W is —(C(R^(wa))(R^(wb)))_(wn)—, Y is —(OCH₂CH₂)_(yn)—O_(yp)—,and Z is —(C(R^(za))(R^(zb)))_(zn);

wherein wn is selected from the group consisting of integers ≥0, and

0 or at least 1 methylene unit of W is independently replaced by -Cyr-,—N(R^(wx))C(O)—, —C(O)N(R^(wx))—, —C(O)—, —OC(O)—, —C(O)O—, —NR^(wx)—,—O—, —S—, —SO—, —SO₂—, —P(R^(wx))—, —P(═O)(R^(wx))—, —N(R^(wx))SO₂—,—SO₂N(R^(wx))—, —C(═S)—, —C(═NR^(wx))—, —N═C— or —C(═N₂)—;

wherein yn is selected from the group consisting of integers ≥0, and ypis 0 or 1;

wherein zn is selected from the group consisting of integers ≥0, and

0 or at least 1 methylene unit of Z is independently replaced by -Cyr-,—N(R^(zx))C(O)—, —C(O)N(R^(zx))—, —C(O)—, —OC(O)—, —C(O)O—, —NR^(zx)—,—O—, —S—, —SO—, —SO₂—, —P(R^(zx))—, —P(═O)(R^(zx))—, —N(R^(zx))SO₂—,—SO₂N(R^(zx))—, (═S)—, —C(═NR^(zx))—, —N═C— or —C(═N₂)—;

-Cyr- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene and 3-10membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or independently substituted with at least 1substituent R^(cx);

wherein each R^(wa), each R^(wb), each R^(za), each R^(zb), each R^(wx),each R^(zx) and each R^(cx) are each independently hydrogen, protium,deuterium, tritium, halogen, —NO₂, —CN, —OR^(r), —SR^(r),—N(R^(ra))(R^(rb)), —C(O)R^(r), —CO₂R^(r), —C(O)C(O)R^(r),—C(O)CH₂C(O)R^(r), —S(O)R^(r), —S(O)₂R^(r), —C(O)N(R^(ra))(R^(rb)),—SO₂N(R^(ra))(R^(rb)), —OC(O)R^(r), —N(R)SO₂R^(r), or a C₁₋₆ aliphaticgroup optionally substituted with R^(r);

wherein each R^(r), each R^(ra) and each R^(rb) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

-L_(b)- represents a peptide residue consisting of 2 to 7 amino acids;

-L_(c)- is selected from the group consisting of:

wherein R^(L1) and R^(L2) are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO₂,—CN, —OH, —SH, —NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H,—S(O)₂H, —C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H and a C₁₋₆ aliphaticgroup;

wherein, X¹ is saturated C, and X¹ is substituted with R^(n);

ring A is selected from the group consisting of: 3-10 membered saturatedor partially unsaturated heterocyclyl and 3-10 membered saturated orpartially unsaturated carbocyclyl, wherein ring A is substituted with 0or at least 1 substituent R^(1a);

when ring A is 3-10 membered saturated or partially unsaturatedcarbocyclyl, ring A is substituted with p L², and L² is not R^(n);

or, when ring A is 3-10 membered saturated or partially unsaturatedheterocyclyl, ring A is substituted with p L²;

L² is —R²-L³-, and R² is used for direct or indirect linking of aligand;

L³ is —(C(R^(3a))(R^(3b)))_(m)—, wherein when L³ comprises a methyleneunit, 0 or at least 1 methylene unit of L³ is independently replaced by—N(R⁴)C(O)—, —C(O)N(R⁴)—, —C(O)—, —OC(O)—, —C(O)O—, —NR⁴—, —O—, —S—,—SO—, —SO₂—, —P(R⁴)—, —P(═O)(R⁴)—, —N(R⁴)SO₂—, —SO₂N(R⁴)—, —C(═S)—,—C(═NR⁴)—, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

R² is selected from the group consisting of: —O—, —(R^(2a))N—, —S— and—P(═O)(R^(2a))—;

L¹ is —(C(R^(5a))(R^(5b)))^(n)—, wherein when L¹ comprises a methyleneunit, 0 or at least 1 methylene unit of 12 is independently replaced by—N(R⁶)C(O)—, —C(O)N(R⁶)—, —C(O)—, —OC(O)—, —C(O)O—, —NR⁶—, —O—, —S—,—SO—, —SO₂—, —P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—, —SO₂N(R⁶)—, —C(═S)—,—C(═NR⁶)—, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

wherein each R^(1a), each R^(2a), each R^(3a), each R^(3b), each R⁴,each R^(5a), each R^(5b), each R⁶ and each R^(n) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R,or a C₁₋₆ aliphatic group optionally substituted with R;

wherein each R, each R^(a) and each R^(b) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

m and n are each independently selected from the group consisting ofintegers ≥0, and p is an integer ≥1.

In one aspect, the present application provides a compound of generalformula (II-F_(x)) or a tautomer, a mesomer, a racemate, an enantiomeror a diastereoisomer thereof, or a mixture thereof, or apharmaceutically acceptable salt thereof,

wherein, L^(x) is L_(ax)-L_(b)-L_(c)-;

L_(ax)- is selected from the group consisting of:

wherein R^(hal) is iodine or bromine;

wherein W is —(C(R^(wa))(R^(wb)))_(wn)—, Y is —(OCH₂CH₂)_(yn)—O_(yp)—,and Z is —(C(R^(za))(R^(zb)))_(zn);

wherein wn is selected from the group consisting of integers ≥0, and

0 or at least 1 methylene unit of W is independently replaced by -Cyr-,—N(R^(wx))C(O)—, —C(O)N(R^(wx))—, —C(O)—, —OC(O)—, —C(O)O—, —NR^(wx)—,—O—, —S—, —SO—, —SO₂—, —P(R^(wx))—, —P(═O)(R^(wx))—, —N(R^(wx))SO₂—,—SO₂N(R^(wx))—, —C(═S)—, —C(═NR^(wx))—, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

wherein yn is selected from the group consisting of integers ≥0, and ypis 0 or 1;

wherein zn is selected from the group consisting of integers ≥0, and

0 or at least 1 methylene unit of Z is independently replaced by -Cyr-,—N(R^(zx))C(O)—, —C(O)N(R^(zx))—, —C(O)—, —OC(O)—, —C(O)O—, —NR^(zx)—,—O—, —S—, —SO—, —SO₂—, —P(R^(zx))—, —P(═O)(R^(zx))—, —N(R^(zx))SO₂—,—SO₂N(R^(zx))—, (═S)—, —C(═NR^(zx))—, —N═N—, —N═C— or —C(═N₂)—;

-Cyr- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene and 3-10membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or independently substituted with at least 1substituent R^(cx);

wherein each R^(wa), each R^(wb), each R^(za), each R^(zb), each R^(wx),each R^(zx) and each R^(cx) are each independently hydrogen, protium,deuterium, tritium, halogen, —NO₂, —CN, —OR^(r), —SR^(r),—N(R^(ra))(R^(rb)), —C(O)R^(r), —CO₂R^(r), —C(O)C(O)R^(r),—C(O)CH₂C(O)R^(r), —S(O)R^(r), —S(O)₂R^(r), —C(O)N(R^(ra))(R^(rb)),—SO₂N(R^(ra))(R^(rb)), —OC(O)R^(r), —N(R)SO₂R^(r), or a C₁₋₆ aliphaticgroup optionally substituted with R^(r);

wherein each R^(r), each R^(ra) and each R^(rb) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

-L_(b)- represents a peptide residue consisting of 2 to 7 amino acids;

-L_(c)- is selected from the group consisting of:

wherein R^(L1) and R^(L2) are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO₂,—CN, —OH, —SH, —NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H,—S(O)₂H, —C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H and a C₁₋₆ aliphaticgroup;

wherein, X¹ is saturated C, and X¹ is substituted with IV;

ring A is selected from the group consisting of: 3-10 membered saturatedor partially unsaturated heterocyclyl and 3-10 membered saturated orpartially unsaturated carbocyclyl, wherein ring A is substituted with 0or at least 1 substituent R^(1a);

when ring A is 3-10 membered saturated or partially unsaturatedcarbocyclyl, ring A is substituted with p L², and L² is not IV;

or, when ring A is 3-10 membered saturated or partially unsaturatedheterocyclyl, ring A is substituted with p L²;

L² is —R²-L³-, and R² is used for direct or indirect linking of aligand;

L³ is —(C(R^(3a))(R^(3b)))_(m)—, wherein when L³ comprises a methyleneunit, 0 or at least 1 methylene unit of L³ is independently replaced by—N(R⁴)C(O)—, —C(O)N(R⁴)—, —C(O)—, —OC(O)—, —C(O)O—, —NR⁴—, —O—, —S—,—SO—, —SO₂—, —P(R⁴)—, —P(═O)(R⁴)—, —N(R⁴)SO₂—, —SO₂N(R⁴)—, —C(═S)—,—C(═NR⁴)—, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

R² is selected from the group consisting of: —O—, —(R^(2′))N—, —S— and—P(═O)(R^(2a))—;

L¹ is —(C(R^(5a))(R^(5b)))_(m)—, wherein when L¹ comprises a methyleneunit, 0 or at least 1 methylene unit of L¹ is independently replaced by—N(R⁶)C(O)—, —C(O)N(R⁶)—, —C(O)—, —OC(O)—, —C(O)O—, —NR⁶—, —O—, —S—,—SO—, —SO₂—, —P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—, —SO₂N(R⁶)—, —C(═S)—,—C(═NR⁶)—, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

wherein each R^(1a), each R^(2a), each R^(3a), each R^(3b), each R⁴,each R^(5a), each R^(5b), each R⁶ and each R^(n) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R,or a C₁₋₆ aliphatic group optionally substituted with R;

wherein each R, each R^(a) and each R^(b) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

m and n are each independently selected from the group consisting ofintegers ≥0, and p is an integer ≥1.

In one aspect, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound comprises the following group ofstructures:

In one aspect, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound comprises a structure shown as formula(III-A):

wherein, R¹ is selected from the group consisting of: —O—, —(R²)N—,—P(═O)(R²)— and —S—;

X is -L¹-CH₂—C(O)—;

L¹ is —(C(R^(3a))(R^(3b)))_(m)—, wherein 0 or at least 1 methylene unitof L¹ is independently replaced by —C(O)—, —C(═S)—, —C(═NR^(4b))— or—C(═N₂)—;

wherein each R², each R^(3a), each R^(3b) and each R^(4b) are eachindependently hydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN,—OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R,—S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R,—N(R)SO₂R, or a C₁₋₆ aliphatic group optionally substituted with R;

wherein each R, each R^(a) and each R^(b) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

m is selected from the group consisting of integers ≥0, and n isselected from the group consisting of integers ≥1;

when R¹ is —O— or —HN—, at least 1 methylene unit of L¹ is independentlyreplaced by —C(O)—, —C(═S)—, —C(═NR^(4b))— or —C(═N₂)—, or each R¹a andeach R^(3b) are not both hydrogen.

In one aspect, the present application provides a compound of generalformula (III-E) or a tautomer, a mesomer, a racemate, an enantiomer or adiastereoisomer thereof, or a mixture thereof, or a pharmaceuticallyacceptable salt thereof,

wherein R¹ is selected from the group consisting of: —O—, —(R²)N—,—P(═O)(R²)— and —S—;

X is -L¹-CH₂—C(O)—;

L¹ is —(C(R^(3a))(R^(3b)))_(m)—, wherein 0 or at least 1 methylene unitof 12 is independently replaced by —C(O)—, —C(═S)—, —C(═NR^(4b))— or—C(═N₂)—;

wherein each R², each R^(3a), each R^(3b) and each R^(4b) are eachindependently hydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN,—OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R,—S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R,—N(R)SO₂R, or a C₁₋₆ aliphatic group optionally substituted with R;

wherein each R, each R^(a) and each R^(b) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

m is selected from the group consisting of integers ≥0, and n isselected from the group consisting of integers ≥1;

when R¹ is —O— or —HN—, at least 1 methylene unit of L¹ is independentlyreplaced by —C(O)—, —C(═S)—, —C(═NR^(4b))— or —C(═N₂)—, or each R^(3a)and each R^(3b) are not both hydrogen.

In one aspect, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound comprises a structure shown as formula(III-C):

wherein, L is -L_(a)-L_(b)-L_(c)-;

-L_(a)- is selected from the group consisting of:

wherein W is —(C(R^(wa))(R^(wb)))_(wn)—, Y is —(OCH₂CH₂)_(yn)—O_(yp),and Z is —(C(R^(za))(R^(zb)))_(zn);

wherein wn is selected from the group consisting of integers ≥0, and

0 or at least 1 methylene unit of W is independently replaced by -Cyr-,—N(R^(wx))C(O)—, —C(O)N(R^(wx))—, —C(O)—, —OC(O)—, —C(O)O—, —N^(Rwx)—,—O—, —S—, —SO—, —SO₂—, —P(R^(wx))—, —P(═O)(R^(wx))—, —N(R^(wx))SO₂—,—SO₂N(R^(wx))—, —C(═S)—, —C(═N^(Rwx))—, —N═C— or —C(═N₂)—;

wherein yn is selected from the group consisting of integers ≥0, and ypis 0 or 1;

wherein zn is selected from the group consisting of integers ≥0, and

0 or at least 1 methylene unit of Z is independently replaced by -Cyr-,—N(R^(zx))C(O)—, —C(O)N(R^(zx))—, —C(O)—, —OC(O)—, —C(O)O—, —NR^(zx)—,—O—, —S—, —SO—, —SO₂—, —P(R^(zx))—, —P(═O)(R^(zx))—, —N(R^(zx))SO₂—,—SO₂N(R^(zx))—, —C(═S)—, —C(═NR^(zx))—, —N═C— or —C(═N₂)—;

-Cyr- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene and 3-10membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or independently substituted with at least 1substituent R^(cx);

wherein each R^(wa), each R^(wb), each R^(za), each R^(zb), each R^(wx),each R^(zx) and each R^(cx) are each independently hydrogen, protium,deuterium, tritium, halogen, —NO₂, —CN, —OR^(r), —SR^(r),—N(R^(ra))(R^(rb)), —C(O)R^(r), —CO₂R^(r), —C(O)C(O)R^(r),—C(O)CH₂C(O)R^(r), —S(O)R^(r), —S(O)₂R^(r), —C(O)N(R^(ra))(R^(rb)),—SO₂N(R^(ra))(R^(rb)), —OC(O)R^(r), —N(R)SO₂R^(r), or a C₁₋₆ aliphaticgroup optionally substituted with R^(r);

wherein each R^(r), each R^(ra) and each R^(rb) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

-L_(b)- represents a peptide residue consisting of 2 to 7 amino acids;

-L_(c)- is selected from the group consisting of:

wherein R^(L1) and R^(L2) are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO₂,—CN, —OH, —SH, —NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H,—S(O)₂H, —C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H and a C₁₋₆ aliphaticgroup;

wherein R¹ is selected from the group consisting of: —O—, —(R²)N—,—P(═O)(R²)— and —S—;

X is -L¹-CH₂—C(O)—;

L¹ is —(C(R^(3a))(R^(3b))_(m)—, wherein 0 or at least 1 methylene unitof 12 is independently replaced by —C(O)—, —C(═S)—, —C(═NR^(4b))— or—C(═N₂)—;

wherein each R², each R^(3a), each R^(3b) and each R^(4b) are eachindependently hydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN,—OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R,—S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R,—N(R)SO₂R, or a C₁₋₆ aliphatic group optionally substituted with R;

wherein each R, each R^(a) and each R^(b) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

m is selected from the group consisting of integers ≥0, and n isselected from the group consisting of integers ≥1;

when R¹ is —O— or —HN—, at least 1 methylene unit of 12 is independentlyreplaced by —C(O)—, —C(═S)—, —C(═NR^(4b))— or —C(═N₂)—, or each R^(3a)and each R^(3b) are not both hydrogen.

In one aspect, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound comprises a structure shown as formula(III-D):

wherein, Ab is a ligand, and an average connection number Na is aninteger or a decimal from 1 to 10;

L is -L_(a)-L_(b)-L_(c)-;

-L_(a)- is selected from the group consisting of:

wherein W is —(C(R^(wa))(R^(wb)))_(wn)—, Y is —(OCH₂CH₂)_(yn)—O_(yp),and Z is —(C(R^(za))(R^(zb)))_(zn);

wherein wn is selected from the group consisting of integers ≥0, and

0 or at least 1 methylene unit of W is independently replaced by -Cyr-,—N(R^(wx))C(O)—, —C(O)N(R^(wx))—, —C(O)—, —OC(O)—, —C(O)O—, —Nltwx-,—O—, —S—, —SO—, —SO₂—, —P(R^(wx))—, —P(═O)(R^(wx))—, —N(R^(wx))SO₂—,—SO₂N(R^(wx))—, —C(═S)—, —C(═NR^(wx))—, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

wherein yn is selected from the group consisting of integers ≥0, and ypis 0 or 1;

wherein zn is selected from the group consisting of integers ≥0, and

0 or at least 1 methylene unit of Z is independently replaced by -Cyr-,—N(R^(zx))C(O)—, —C(O)N(R^(zx))—, —C(O)—, —OC(O)—, —C(O)O—, —NR′—, —O—,—S—, —SO—, —SO₂—, —P(R^(zx))—, —P(═O)(R^(zx))—, —N(R^(zx))SO₂—,—SO₂N(R^(zx))—, (═S)—, —C(═NR^(zx))—, —N═N—, —N═C— or —C(═N₂)—;

-Cyr- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene and 3-10membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or independently substituted with at least 1substituent R^(cx);

wherein each R^(wa), each R^(wb), each R^(za), each R^(zb), each R^(wx),each R^(zx) and each R^(cx) are each independently hydrogen, protium,deuterium, tritium, halogen, —NO₂, —CN, —OR^(r), —SR^(r),—N(R^(ra))(R^(rb)), —C(O)R^(r), —CO₂R^(r), —C(O)C(O)R^(r),—C(O)CH₂C(O)R^(r), —S(O)R^(r), —S(O)₂R^(r), —C(O)N(R^(ra))(R^(rb)),—SO₂N(R^(ra))(R^(rb)), —OC(O)R^(r), —N(R)SO₂R^(r), or a C₁₋₆ aliphaticgroup optionally substituted with R^(r);

wherein each R^(r), each R_(ra) and each R^(ra) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

-L_(b)- represents a peptide residue consisting of 2 to 7 amino acids;

-L_(c)- is selected from the group consisting of:

wherein R^(L1) and R^(L2) are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO₂,—CN, —OH, —SH, —NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H,—S(O)₂H, —C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H and a C₁₋₆ aliphaticgroup;

wherein R¹ is selected from the group consisting of: —O—, —(R²)N—,—P(═O)(R²)— and —S—;

X is -L¹-CH₂—C(O)—;

L¹ is —(C(R^(3a))(R^(3b)))_(m)—, wherein 0 or at least 1 methylene unitof 12 is independently replaced by —C(O)—, —C(═S)—, —C(═NR^(4b))— or—C(═N₂)—;

wherein each R², each R^(3a), each R^(3b) and each R^(4b) are eachindependently hydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN,—OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R,—S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R,—N(R)SO₂R, or a C₁₋₆ aliphatic group optionally substituted with R;

wherein each R, each IV and each R^(b) are each independently hydrogen,protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH, —NH₂, —C(O)H,—CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H, —C(O)NH₂, —SO₂NH₂,—OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

m is selected from the group consisting of integers ≥0, and n isselected from the group consisting of integers ≥1;

when R¹ is —O— or —HN—, at least 1 methylene unit of 12 is independentlyreplaced by —C(O)—, —C(═S)—, —C(═NR^(4b))— or —C(═N₂)—, or each R^(3a)and each R^(3b) are not both hydrogen.

In one aspect, the present application provides a compound of generalformula (III-F) or a tautomer, a mesomer, a racemate, an enantiomer or adiastereoisomer thereof, or a mixture thereof, or a pharmaceuticallyacceptable salt thereof,

wherein, L^(x) is L_(ax)-L_(b)-L_(c)-;

L_(ax)- is selected from the group consisting of:

wherein R^(hal) is iodine or bromine;

wherein W is —(C(R^(wa))(R^(wb)))_(wn)—, Y is —(OCH₂CH₂)_(yn)—O_(yp),and Z is —(C(R^(za))(R^(zb)))_(zn);

wherein wn is selected from the group consisting of integers ≥0, and

0 or at least 1 methylene unit of W is independently replaced by -Cyr-,—N(R^(wx))C(O)—, —C(O)N(R^(wx))—, —C(O)—, —OC(O)—, —C(O)O—, —NR^(wx)—,—O—, —S—, —SO—, —SO₂—, —P(R^(wx))—, —P(═O)(R^(wx))—, —N(R^(wx))SO₂—,—SO₂N(R^(wx))—, —C(═S)—, —C(═NR^(wx))—, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

wherein yn is selected from the group consisting of integers ≥0, and ypis 0 or 1;

wherein zn is selected from the group consisting of integers ≥0, and

0 or at least 1 methylene unit of Z is independently replaced by -Cyr-,—N(R^(zx))C(O)—, —C(O)N(R″)—, —C(O)—, —OC(O)—, —C(O)O—, —NR′—, —O—, —S—,—SO—, —SO₂—, —P(R″)—, —P(═O)(R^(zx))—, —N(R^(zx))SO₂—, —SO₂N(R′)—,(═S)—, —C(═NR^(zx))—, —N═N—, —N═C— or —C(═N₂)—;

-Cyr- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene and 3-10membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or independently substituted with at least 1substituent R^(cx);

wherein each R^(wa), each R^(wb), each R^(za), each R^(zb), each R^(wx),each R^(zx) and each R^(cx) are each independently hydrogen, protium,deuterium, tritium, halogen, —NO₂, —CN, —OR^(r), —SR^(r),—N(R^(ra))(R^(rb)), —C(O)R^(r), —CO₂R^(r), —C(O)C(O)R^(r),—C(O)CH₂C(O)R^(r), —S(O)R^(r), —S(O)₂R^(r), —C(O)N(R^(ra))(R^(rb)),—SO₂N(R^(ra))(R^(rb)), —OC(O)R^(r), —N(R)SO₂R^(r), or a C₁₋₆ aliphaticgroup optionally substituted with R^(r);

wherein each R^(r), each R^(ra) and each R^(rb) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

-L_(b)- represents a peptide residue consisting of 2 to 7 amino acids;

-L_(c)- is selected from the group consisting of:

wherein R^(L1) and R^(L2) are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO₂,—CN, —OH, —SH, —NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H,—S(O)₂H, —C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H and a C₁₋₆ aliphaticgroup;

wherein R¹ is selected from the group consisting of: —O—, —(R²)N—,—P(═O)(R²)— and —S—;

X is -L¹-CH₂—C(O)—;

L¹ is —(C(R^(3a))(R^(3b)))_(m)—, wherein 0 or at least 1 methylene unitof 12 is independently replaced by —C(O)—, —C(═S)—, —C(═NR^(4b))— or—C(═N₂)—;

wherein each R², each R^(1a), each R^(3b) and each R^(4b) are eachindependently hydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN,—OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R,—S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R,—N(R)SO₂R, or a C₁₋₆ aliphatic group optionally substituted with R;

wherein each R, each R^(a) and each R^(b) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

m is selected from the group consisting of integers ≥0, and n isselected from the group consisting of integers ≥1;

when R¹ is —O— or —HN—, at least 1 methylene unit of 12 is independentlyreplaced by —C(O)—, —C(═S)—, —C(═NR^(4b))— or —C(═N₂)—, or each R^(3a)and each R^(3b) are not both hydrogen.

In one aspect, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the ligand-drug conjugate comprises the following groupof structures:

In one aspect, the present application provides a method for preparingthe compound or the tautomer, the mesomer, the racemate, the enantiomeror the diastereoisomer thereof, or the mixture thereof, or thepharmaceutically acceptable salt thereof disclosed herein, whichcomprises contacting a ligand Ab with the structure shown as formula(II-FX) disclosed herein.

In one aspect, the present application provides a method for preparingthe compound or the tautomer, the mesomer, the racemate, the enantiomeror the diastereoisomer thereof, or the mixture thereof, or thepharmaceutically acceptable salt thereof disclosed herein, whichcomprises contacting a ligand Ab with the structure shown as formula(IMF) disclosed herein.

In one aspect, the present application provides a pharmaceuticalcomposition, which comprises the compound or the tautomer, the mesomer,the racemate, the enantiomer or the diastereoisomer thereof, or themixture thereof, or the pharmaceutically acceptable salt thereofdisclosed herein, and a pharmaceutically acceptable carrier.

In one aspect, the present application provides use of the compound orthe tautomer, the mesomer, the racemate, the enantiomer or thediastereoisomer thereof, or the mixture thereof, or the pharmaceuticallyacceptable salt thereof disclosed herein, and/or the pharmaceuticalcomposition disclosed herein, in preparing a medicament for treatingand/or preventing a tumor.

Other aspects and advantages of the present application will be readilyapparent to those skilled in the art from the following detaileddescription. Only exemplary embodiments of the present application havebeen shown and described in the following detailed description. As thoseskilled in the art will recognize, the content of the presentapplication enables those skilled in the art to make changes to thespecific embodiments disclosed without departing from the spirit andscope of the invention to which the present application pertains.Accordingly, descriptions in the drawings and specification are onlyillustrative rather than restrictive.

BRIEF DESCRIPTION OF THE DRAWING

Specific features of the invention to which the present applicationpertains are set forth in appended claims. Features and advantages ofthe invention to which the present application pertains will be betterunderstood by reference to the exemplary embodiments and drawingsdescribed in detail below. The drawings are briefly described asfollows:

FIGS. 1-16 are graphs showing in vivo tumor inhibition results of thecompounds disclosed herein.

DETAILED DESCRIPTION

The embodiments of the present invention are described below withreference to specific examples, and other advantages and effects of thepresent invention will be readily apparent to those skilled in the artfrom the disclosure of the present specification.

Definitions of Terms

In the present application, the term “ligand” generally refers to amacromolecular compound capable of recognizing and binding to an antigenor receptor associated with a target cell. The role of ligands may be topresent the drug to a target cell population to which the ligand binds,and the ligands include, but are not limited to, protein hormones,lectin, growth factors, antibodies, or other molecules capable ofbinding to a cell, a receptor and/or an antigen. In the presentapplication, the ligand may be represented as Ab, the ligand antigenforms a linking bond with the linking unit through a heteroatom on theligand, and the ligand may be an antibody or an antigen-binding fragmentthereof, wherein the antibody may be selected from the group consistingof a chimeric antibody, a humanized antibody, a fully human antibody ora murine antibody, and the antibody may be a monoclonal antibody. Forexample, the antibody may be an antibody that targets the followingtarget points: HER2, HER3, B7H3, TROP2, Claudin 18.2, CD30, CD33, CD70or EGFR. For example, the antibody may be an antibody that targets thefollowing target points: 5T4, AGS-16, ANGPTL4, ApoE, CD19, CTGF, CXCR5,FGF2, MCPT8, MFI2, MS4A7, NCA, Sema5b, SLITRK6, STC2, TGF, 0772P, 5T4,ACTA2, ADGRE1, AG-7, AIF1, AKR1C1, AKR1C2, ASLG659, Axl, B7H3, BAFF-R,BCMA, BMPR1B, BNIP3, C1QA, C1QB, CA6, CADM1, CCD79b, CCL5, CCR5, CCR7,CD11c, CD123, CD138, CD142, CD147, CD166, CD19, CD19, CD22, CD21, CD20,CD205, CD22, CD223, CD228, CD25, CD30, CD33, CD37, CD38, CD40, CD45,CD45 (PTPRC), CD46, CD47, CD49D (ITGA4), CD56, CD66e, CD70, CD71, CD72,CD74, CD79a, CD79b, CD80, CDCP1, CDH11, CD11b, CEA, CEACAM5, c-Met,COL6A3, COL7A1, CRIPTO, CSF1R, CTSD, CTSS, CXCL11, CXCL10, DDIT4, DLL3,DLL4, DR5, E16, EFNA4, EGFR, EGFRvIII, EGLN, EGLN3, EMR2, ENPP3, EpCAM,EphA2, EphB2R, ETBR, FcRH2, FcRH1, FGFR2, FGFR3, FLT3, FOLR-α, GD2,GEDA, GPC-1, GPNMB, GPR20, GZMB, HER2, HER3, HLA-DOB, HMOX1, IFI6, IFNG,IGF-1R, IGFBP3, IL10RA1, IL-13R, IL-2, IL20Ra, IL-3, IL-4, IL-6, IRTA2,KISS1R, KRT33A, LIV-1, LOX, LRP-1, LRRC15, LUM, LY64, LY6E, Ly86, LYPD3,MDP, MMP10, MMP14, MMP16, MPF, MSG783, MSLN, MUC-1, NaPi2b, Napi3b,Nectin-4, Nectin-4, NOG, P2X5, pCAD, P-Cadherin, PDGFRA, PDK1, PD-L1,PFKFB3, PGF, PGK1, PIK3AP1, PIK3CD, PLOD2, PSCA, PSCAhlg, PSMA, PSMA,PTK7, P-Cadherin, RNF43, NaPi2b, ROR1, ROR2, SERPINE1, SLC39A6, SLTRK6,STAT1, STEAP1, STEAP2, TCF4, TENB2, TGFB1, TGFB2, TGFBR1, TNFRSF21,TNFSF9, Trop-2, TrpM4, Tyrol, UPK1B, VEGFA, WNT5A, epidermal growthfactors, brevican, mesothelin, sodium phosphate cotransporter 2B,Claudin 18.2, endothelin receptors, mucins (such as mucin 1 and mucin16), guanylate cyclase C, integrin a4p7, integrin a5p6, trophoblastglycoprotein, or tissue factors.

In the present application, the term “cytotoxic drug” generally refersto a toxic drug, and the cytotoxic drug may have a chemical moleculewithin the tumor cell that is strong enough to disrupt its normalgrowth. Cytotoxic drugs can kill tumor cells at a sufficiently highconcentration. The “cytotoxic drug” may include toxins, such as smallmolecule toxins or enzymatically active toxins of bacterial, fungal,plant or animal origin, radioisotopes (e.g., At²¹¹, I¹³¹, I¹²⁵, Y⁹⁰,Re¹⁸⁶, Re¹⁸⁸, Sm¹⁵³, Bi²¹², P³² or radioactive isotopes of Lu), toxicdrugs, chemotherapeutic drugs, antibiotics and nucleolytic enzymes; forexample, the cytotoxic drug may be toxic drugs, including but notlimited to camptothecin derivatives, which, for example, may be thecamptothecin derivative exatecan (chemical name: (1S,9S)-1-amino-9-ethyl-5-fluoro-2,3-dihydro-9-hydroxy-4-methyl-1H,12H-benzo[de]pyrano[3′,4′:6,7]imidazo[1,2-b]quinoline-10,13 (9H,15H)-dione).

In the present application, the term “linker structure” generally refersto a chemical structural fragment or bond, which is linked to a ligandat one end and linked to a cytotoxic drug at the other end, or linked toother linkers and then linked to the cytotoxic drug. The direct orindirect linking of a ligand may mean that the group is directly linkedto the ligand via a covalent bond, and may also be linked to the ligandvia a linker structure. For example, the linker structure may be astructure shown as -L_(a)x-L_(b)-L_(c)- and/or -L_(a)-L_(b)-L_(c)-described herein. For example, a chemical structure fragment or bondcomprising an acid-labile linker structure (e.g., hydrazone), aprotease-sensitive (e.g., peptidase-sensitive) linker structure, aphotolabile linker structure, a dimethyl linker structure or adisulfide-containing linker structure may be used as a linker structure.

In the present application, the term a structure being “optionallylinked to other molecular moieties” generally means that the structureis not linked to any other chemical structure, or that the structure islinked (e.g., via a chemical bond or a linker structure) to one or moreother chemical structures (e.g., ligands described herein) differentfrom the structure.

In the present application, the term “ligand-drug conjugate” generallymeans that a ligand is linked to a biologically active cytotoxic drugvia a stable linking unit. In the present application, the “ligand-drugconjugate” may be an antibody-drug conjugate (ADC), which may mean thata monoclonal antibody or an antibody fragment is linked to abiologically active cytotoxic drug via a stable linking unit.

In the present application, the term “antibody or antigen-bindingfragment thereof” generally refers to that immunological bindingreagents extend to all antibodies from all species, including dimeric,trimeric and multimeric antibodies; bispecific antibodies; chimericantibodies; fully humanized antibodies; humanized antibodies;recombinant and engineered antibodies and fragments thereof. The term“antibody or fragment thereof” may refer to any antibody-like moleculehaving an antigen-binding region, and includes small molecule fragments,such as Fab′, Fab, F(ab)₂, single domain antibodies (DABs), Fv, scFv(single chain Fv), linear antibodies, and diabodies. The term“antigen-binding fragment” may refer to one or more fragments of anantibody that retain the ability to specifically bind to an antigen. Forexample, a fragment of a full-length antibody can be used to perform theantigen-binding function of the antibody. Techniques for preparing andusing various antibody-based constructs and fragments are well known inthe art. The antibody may include one or more of an anti-HER2(ErbB2)antibody, an anti-EGFR antibody, an anti-B7-H3 antibody, an anti-c-Metantibody, an anti-HER3(ErbB3) antibody, an anti-HER4(ErbB4) antibody, ananti-CD20 antibody, an anti-CD22 antibody, an anti-CD30 antibody, ananti-CD33 antibody, an anti-CD44 antibody, an anti-CD56 antibody, ananti-CD70 antibody, an anti-CD73 antibody, an anti-CD105 antibody, ananti-CEA antibody, an anti-A33 antibody, an anti-Cripto antibody, ananti-EphA2 antibody, an anti-G250 antibody, an anti-MUC1 antibody, ananti-Lewis Y antibody, an anti-TROP2 antibody, an anti-Claudin 18.2antibody, an anti-VEGFR antibody, an anti-GPNMB antibody, ananti-Integrin antibody, an anti-PSMA antibody, an anti-Tenascin-Cantibody, an anti-SLC44A4 antibody and an anti-Mesothelin antibody; forexample, it may be trastuzumab or pertuzumab.

In the present application, the term “chimeric antibody” generallyrefers to an antibody obtained by fusing a variable region of a murineantibody and a constant region of a human antibody, which can reduce animmune response induced by the murine antibody. For establishment of achimeric antibody, a hybridoma secreting murine specific monoclonalantibody can be established, and a variable region gene is cloned fromthe mouse hybridoma cells; then a constant region gene of human antibodycan be cloned as required, and the mouse variable region gene and thehuman constant region gene are connected to form a chimeric gene; thenthe chimeric gene is inserted into an expression vector, whereinchimeric antibody molecules can be expressed in a eukaryotic system or aprokaryotic system.

In the present application, the term “humanized antibody”, also referredto as CDR-grafted antibody, generally refers to an antibody produced bygrafting mouse CDR sequences into a human antibody variable regionframework, i.e., an antibody produced in a different type of humangermline antibody framework sequence. Therefore, the heterogeneousreaction induced by the presence of a large number of mouse proteincomponents in the chimeric antibody can be overcome. Such frameworksequences can be obtained from public DNA databases or disclosedreferences that include germline antibody gene sequences. For example,germline DNA sequences of human heavy and light chain variable regiongenes can be obtained from the “VBase” human germline sequence database.

In the present application, the term “fully humanized antibody”, “fullyhuman antibody” or “completely human antibody”, also known as “fullyhumanized monoclonal antibody”, may have both humanized variable regionand constant region so as to eliminate immunogenicity and toxic sideeffects. The development of monoclonal antibodies has four stages,namely murine monoclonal antibodies, chimeric monoclonal antibodies,humanized monoclonal antibodies and fully humanized monoclonalantibodies. The antibodies or ligands described herein can be fullyhumanized monoclonal antibodies. Relevant technologies for thepreparation of fully human antibodies may be: human hybridomatechnology, EBV-transformed B-lymphocyte technology, phage displaytechnology, transgenic mouse antibody preparation technology, singleB-cell antibody preparation technology, and the like.

In the present application, the term “CDR” generally refers to one ofthe 6 hypervariable regions within the variable domain of an antibodywhich contribute primarily to antigen binding. One of the most commondefinitions of the 6 CDRs is provided by Kabat E. A. et al., (1991)Sequences of proteins of immunological interest. NIH Publication91-3242; Chothia et al., “Canonical Structures For the HypervariableRegions of Immunoglobulins”, J. Mol. Biol.196: 901 (1987); and MacCallumet al., “Antibody-Antigen Interactions: Contact Analysis and BindingSite Topography”, J. Mol. Biol. 262:732 (1996). As used herein, theKabat definition of CDRs can be applied to CDR1, CDR2 and CDR3 of thelight chain variable domain (CDR L1, CDR L2, CDR L3 or L1, L2, L3), andCDR1, CDR2 and CDR3 of the heavy chain variable domain (CDR H1, CDR H2,CDR H3 or H1, H2, H3).

In the present application, the term “methylene” generally refers to aresidue derived by removal of two hydrogen atoms from a group having 1carbon atom. Methylene may be substituted or unsubstituted, or replacedor unreplaced. The term “alkylene” generally refers to a saturatedlinear or branched aliphatic hydrocarbon group having 2 residues derivedfrom the parent alkane by removal of two hydrogen atoms from the samecarbon atom or two different carbon atoms, and it may be a linear orbranched group containing 1 to 20 carbon atoms, such as alkylenecontaining 1 to 12 carbon atoms (e.g., 1 to 6 carbon atoms).Non-limiting examples of alkylene groups include, but are not limitedto, methylene(—CH₂—), 1,1-ethylidene(—CH(CH₃)—),1,2-ethylidene(—CH₂CH₂)—, 1,1-propylidene(—CH(CH₂CH₃)—),1,2-propylidene(—CH₂CH(CH₃)—), 1,3-propylidene(—CH₂CH₂CH₂—),1,4-butylidene(-CH₂CH₂CH₂CH₂—), 1,5-butylidene(—CH₂CH₂CH₂CH₂CH₂—), andthe like. Alkylene groups may be substituted or unsubstituted, replacedor unreplaced. For example, when it is substituted, substitution with asubstituent may be performed at any available linking point, and thesubstituent is preferably independently optionally selected from one ormore of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen,mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl,heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio,heterocycloalkylthio, and oxo, and it may, e.g., be hydrogen, protium,deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH, —NH₂, —C(O)H, —CO₂H,—C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H, —C(O)NH₂, —SO₂NH₂, —OC(O)H,—N(H)SO₂H or a C₁₋₆ aliphatic group.

In the present application, the term “arylene” generally refers to agroup having two residues derived by removal of two hydrogen atoms fromthe same carbon atom or two different carbon atoms of the aromatic ring.The term “aromatic ring” may refer to a 6-14 membered all-carbonmonocyclic ring or fused polycyclic ring (i.e., rings which shareadjacent pairs of carbon atoms) having a conjugated 7c-electron system,and it may be 6-10 membered, such as benzene and naphthalene. Thearomatic ring can be fused to a heteroaryl, heterocyclyl or cycloalkylring, wherein the ring connected to the parent moiety is the aryl ring.Aryl may be substituted or unsubstituted, and when it is substituted,the substituent is preferably one or more of the following groupsindependently selected from the group consisting of: alkyl, alkenyl,alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy,nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl,cycloalkoxy, heterocycloalkoxy, cycloalkylthio and heterocycloalkylthio.

In the present application, the term “heteroarylene” generally refers toa group having two residues derived by removal of two hydrogen atomsfrom the same carbon atom or two different carbon atoms of theheteroaromatic ring. The term “heteroaromatic ring” refers to aheteroaromatic system comprising 1 to 4 heteroatoms and 5 to 14 ringatoms, wherein the heteroatoms may be selected from the group consistingof: oxygen, sulfur and nitrogen. Heteroaryl may be 5-10 membered and maybe 5- or 6-membered, such as furanyl, thienyl, pyridinyl, pyrrolyl,N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl and tetrazolyl. Theheteroaromatic ring can be fused to an aryl, heterocyclyl or cycloalkylring, wherein the ring connected to the parent moiety is theheteroaromatic ring. Heteroarylene may be optionally substituted orunsubstituted, and when it is substituted, the substituent is preferablyone or more of the following groups independently selected from thegroup consisting of: alkyl, alkenyl, alkynyl, alkoxy, alkylthio,alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl,heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy,cycloalkylthio and heterocycloalkylthio.

In the present application, the term “heterocyclylene” generally refersto a 3-7 membered monocyclic structure, a fused 7-10 membered bicyclicheterocyclic structure or a bridged 6-10 membered bicyclic heterocyclicstructure that is stable and non-aromatic. These cyclic structures maybe saturated or partially saturated, and contain one or more heteroatomsin addition to carbon atoms, wherein the heteroatoms may be selectedfrom the group consisting of: oxygen, sulfur and nitrogen. For example,they contain 1 to 4 heteroatoms as defined above. When used to refer toatoms on a heterocyclic cyclic structure, the term “nitrogen” mayinclude nitrogen that undergoes a substitution reaction. Heterocyclylenemay be substituted or unsubstituted.

In the present application, the term “carbocyclylene” generally refersto a group having two residues derived by removal of two hydrogen atomsfrom the same carbon atom or two different carbon atoms of the carbonring. The term “carbon ring” generally refers to a saturated orpartially unsaturated monocyclic or polycyclic cyclic hydrocarbon, andit contains 3 to 20 carbon atoms, may contain 3 to 12 carbon atoms, maycontain 3 to 10 carbon atoms, and may contain 3 to 8 carbon atoms.Non-limiting examples of monocyclic carbon ring include cyclopropane,cyclobutane, cyclopentane, cy cl op entene, cyclohexane, cyclohexene,cyclohexadiene, cycloheptane, cycloheptatriene, cyclooctane, and thelike; polycyclic carbon ring may include spiro, fused and bridged carbonrings. Carbocyclylene may be substituted or unsubstituted.

In the present application, the term “partially unsaturated” generallymeans that the cyclic structure contains at least one double or triplebond between the ring molecules. The term “partially unsaturated”encompasses cyclic structures having multiple sites of unsaturation, butis not intended to include aromatic or heteroaromatic rings definedherein. The term “unsaturated” means that the moiety has one or moredegrees of unsaturation.

In the present application, the term “halogen” generally refers tofluorine, chlorine, bromine or iodine, and it may be, for example,fluorine or chlorine.

In the present application, the term “aliphatic group” generally refersto a linear hydrocarbon, branched hydrocarbon or cyclic hydrocarbonhaving 1 to 12 carbon atoms, and the hydrocarbon may be either a fullysaturated hydrocarbon or a hydrocarbon with one or more unsaturatedunits, but the unsaturated units are not aromatic groups. For example,suitable aliphatic groups may include substituted or unsubstitutedlinear, branched or cyclic alkyl, alkenyl, alkynyl and mixtures thereof,such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.For example, aliphatic groups have 1 to 12, 1 to 8, 1 to 6, 1 to 4 or 1to 3 carbon atoms.

In the present application, the term “optional” or “optionally”generally means that the event or circumstance subsequently describedmay, but not necessarily, occur, and that the description includesinstances where the event or circumstance occurs or does not occur. Forexample, “heterocyclyl group optionally substituted with alkyl” meansthat alkyl may be, but not necessarily, present, and that thedescription may include instances where the heterocyclyl group is or isnot substituted with alkyl.

In the present application, the term “substituted” generally means thatone or more hydrogen atoms in the group, for example, up to 5 (e.g., 1to 3) hydrogen atoms, are each independently substituted with acorresponding number of substituents. A substituent is only in itspossible chemical position, and those skilled in the art will be able todetermine (by experiments or theories) possible or impossiblesubstitution without undue efforts. For example, it may be unstable whenamino or hydroxy having a free hydrogen is bound to a carbon atom havingan unsaturated (such as olefin) bond.

In the present application, the term “0 or more (e.g., 0 or at least 1,0 or 1, or 0) methylene units are replaced “generally means that whenthe structure comprises one or more methylene units, the one or moremethylene units may be unsubstituted or replaced by one or more groupsthat are not methylene (e.g., —NHC(O)—, —C(O)NH—, —C(O)—, —OC(O)—,—C(O)O—, —NH—, —O—, —S—, —SO—, —SO₂—, —PH—, —P(═O)H—, —NHSO2-, —SO₂NH—,—C(═S)—, —C(═NH)—, —N═N—, —C═N—, —N═C— or —C(═N₂)—).

One or more hydrogen atoms in the group, for example, up to 5 (e.g., 1to 3) hydrogen atoms, are each independently substituted with acorresponding number of substituents. A substituent is only in itspossible chemical position, and those skilled in the art will be able todetermine (by experiments or theories) possible or impossiblesubstitution without undue efforts. For example, it may be unstable whenamino or hydroxy having a free hydrogen is bound to a carbon atom havingan unsaturated (such as olefin) bond.

In the present application, the term “compound” generally refers to asubstance having two or more different elements. For example, thecompound disclosed herein may be an organic compound. For example, thecompound disclosed herein may be a compound having a molecular weight ofno more than 500, a compound having a molecular weight of no more than1000, a compound having a molecular weight of no less than 1000, or acompound having a molecular weight of no less than 10,000 or no lessthan 100,000. In the present application, the compound may also refer toa compound that involves linking by a chemical bond, for example, acompound where one or more molecules having a molecular weight of nomore than 1000 are linked, by a chemical bond, to a biologicalmacromolecule, wherein the biological macromolecule may bepolysaccharide, protein, nucleic acid, polypeptide, and the like. Forexample, the compound disclosed herein may include a compound where aprotein is linked to one or more molecules having a molecular weight ofno more than 1000, may include a compound where a protein is linked toone or more molecules having a molecular weight of no more than 10,000,and may include a compound where a protein is linked to one or moremolecules having a molecular weight of no more than 100,000.

In some embodiments, the cytotoxic drug of the compound disclosed hereinis directly or indirectly linked to a ligand. In some embodiments, thecytotoxic drug of the compound disclosed herein is directly linked to aligand via a covalent bond. In some embodiments, the cytotoxic drug ofthe compound disclosed herein is linked to a ligand via a linkerstructure. In some embodiments, the compound disclosed herein is aligand-drug conjugate having a ligand linked to a cytotoxic drug via alinker structure, wherein the cytotoxic drug comprises the structuralformula II-A, II-A-1, II-A-2, II-A-3, II-A-4, II-A-5, II-A-6, II-A-7,II-A-8, II-A-9, II-A-10, II-A-11 or II-A-12 disclosed herein, whereinring A, X¹, L¹, and R² are each defined as in embodiments of the firstaspect.

In some embodiments, the compound disclosed herein is a ligand-drugconjugate or ADC. In some embodiments, a “mixture” of a compound refersto a composition comprising one or more of the compound and thetautomer, the mesomer, the racemate, the enantiomer and thediastereoisomer thereof. In some embodiments, a “mixture” of a compoundrefers to a composition comprising its heterogeneous DAR distribution.In one embodiment, a mixture of a compound comprises an ADC having a DARdistribution of 1 to 10, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 (a drugloading of 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10). In one embodiment, the ADCmixture comprises the following two: an ADC with DAR of 6 or less (adrug loading of 6 or less) and an ADC with DAR of 8 or more (a drugloading of 8 or more).

Unless otherwise indicated, the structures described herein may alsoinclude compounds that differ only in the presence or absence of one ormore isotopically enriched atoms. For example, compounds having astructure identical to the structure disclosed herein except for thesubstitution of the hydrogen atom with deuterium or tritium or thesubstitution of the carbon atom with carbon 13 or carbon 14 are withinthe scope of the present application.

In the present application, the term “pharmaceutical composition”generally refers to a mixture containing one or more of the compoundsdescribed herein or a physiologically/pharmaceutically acceptable saltor pro-drug thereof, and other chemical components, for examplephysiologically/pharmaceutically acceptable carriers and excipients. Thepharmaceutical composition may promote the administration to anorganism, which facilitates the absorption of the active ingredient,thereby exerting biological activities. For preparation of conventionalpharmaceutical compositions, reference can be made to ChinesePharmacopoeia.

In the present application, the term “pharmaceutically acceptable salt”generally refers to a salt of a compound or ligand-drug conjugatedisclosed herein, or a salt of a compound described herein. Such saltsmay be safe and/or effective when used in a mammals and may possess therequired biological activity, and the antibody-antibody drug conjugatecompound disclosed herein may form a salt with an acid, and non-limitingexamples of pharmaceutically acceptable salts include: hydrochloride,hydrobromide, hydriodate, sulphate, bisulfate, citrate, acetate,succinate, ascorbate, oxalate, nitrate, sorbate, hydrophosphate,dihydrophosphate, salicylate, hydrocitrate, tartrate, maleate, fumarate,formate, benzoate, mesylate, ethanesulfonate, benzenesulphonate andp-toluenesulfonate.

In the present application, the term “solvate” or “solvent compound”generally refers to a pharmaceutical acceptable solvate formed by aligand-drug conjugate compound disclosed herein and one or more solventmolecules, and non-limiting examples of solvent molecules include water,ethanol, acetonitrile, isopropanol, DMSO and ethyl acetate.

The term “drug loading” generally refers to the average amount ofcytotoxic drug loaded per ligand and may also be expressed as the ratioof cytotoxic drug to antibody, and the cytotoxic drug loading may rangefrom 0 to 12 (e.g., 1 to 10) cytotoxic drugs per ligand (Ab). In theembodiments of the present application, the drug loading is expressed asN^(a), and exemplary values may be an average of 1, 2, 3, 4, 5, 6, 7, 8,9 and 10. The drug loading per ADC molecule after the coupling reactioncan be characterized by conventional methods such as UV/visiblespectroscopy, mass spectrometry, ELISA assays and HPLC.

The pharmaceutical composition may be in the form of a sterileinjectable aqueous or oily suspension for intramuscular and subcutaneousadministration. The suspension can be prepared according to a knowntechnique using those suitable dispersing agents or wetting agents andsuspending agents described above. The sterile injectable formulationmay also be a sterile injection or suspension prepared in a parenterallyacceptable non-toxic diluent or solvent, such as a solution prepared in1,3-butanediol. In addition, a sterile fixed oil may be convenientlyused as a solvent or a suspending medium. For example, any blend fixedoil including synthetic mono- or di-glycerides can be used. In addition,fatty acids such as oleic acid may also be used in the preparation ofinjections.

In the present application, the term “comprise” “comprising”, “contain”or “containing” is generally intended to include the explicitlyspecified features without excluding other elements. The terms “no lessthan” and “no more than” generally refer to the situations where thenumber itself is included.

In the present application, the term “about” generally means varying by0.5%-10% above or below the stated value, for example, varying by 0.5%,1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%,8%, 8.5%, 9%, 9.5% or 10% above or below the stated value.

DETAILED DESCRIPTION OF THE INVENTION

In a first aspect, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(I-A):

wherein, R¹ may be selected from the group consisting of: —O—, —(R²)N—,—P(═O)(R²)— and —S—;

L² may be —(C(R^(3a))(R^(3b)))_(m)—R, and m may be selected from thegroup consisting of integers ≥1;

wherein 0 or no less than 1 methylene unit of L² may be independentlyreplaced by -Cy-, —N(R⁴)C(O)—, —C(O)N(R⁴)—, —C(O)—, —OC(O)—, —C(O)O—,—NR⁴—, —O—, —S—, —SO—, —SO₂—, —P(R⁴)—, —P(═O)(R⁴)—, —N(R⁴)SO₂—,—SO₂N(R⁴)—, —C(═S)—, —C(═NR⁴)—, —N═C— or —C(═N₂)—;

L¹ may be —(C(R^(5a))(R^(5b)))_(n)—, and n may be selected from thegroup consisting of integers ≥1;

wherein 0 or no less than 1 methylene unit of L¹ may be independentlyreplaced by -Cy-, —N(R⁶)C(O)—, —C(O)N(R⁶)—, —C(O)—, —OC(O)—, —C(O)O—,—NR⁶—, —O—, —S—, —SO—, —SO₂—, —P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—,—SO₂N(R⁶)—, —C(═S)—, —C(═NR⁶)—, —N═C— or —C(═N₂)—;

-Cy- may be selected from the group consisting of: 6-10 memberedarylene, 5-8 membered heteroarylene, 3-10 membered heterocyclylene, and3-10 membered saturated or partially unsaturated carbocyclylene, wherein-Cy- is unsubstituted or may be independently substituted with no lessthan 1 substituent R⁷;

wherein each R^(3a), each R^(3b), each R⁴, each R^(5a), each R^(5b) andeach R⁶ may each independently be hydrogen, protium, deuterium, tritium,halogen, —NO₂, —CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R,—C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group whichmay be optionally substituted with R; or, R^(3a) and R^(5a), R⁴ andR^(5a), R^(3a) and R⁶ or R⁴ and R⁶ may each independently optionallyform a ring B together with an atom therebetween, wherein the ring B maybe selected from the group consisting of: 5-8 membered heteroarylene and3-10 membered saturated or partially unsaturated heterocyclylene, andthe ring B is unsubstituted or may be substituted with no less than 1substituent R⁸;

wherein each R², each R⁷ and each R⁸ may each independently be hydrogen,protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R,or a C₁₋₆ aliphatic group which may be optionally substituted with R;

wherein each R, each R^(a) and each R^(b) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(I-A):

wherein, R¹ may be selected from the group consisting of: —O—, —(R²)N—,—P(═O)(R²)— and —S—;

L² may be —(C(R^(3a))(R^(3b)))_(m)—R, and m may be selected from thegroup consisting of integers ≥1;

wherein 0 or no less than 1 methylene unit of L² may be independentlyreplaced by -Cy-, —N(R⁴)C(O)—, —C(O)N(R⁴)—, —C(O)—, —OC(O)—, —C(O)O—,—NR⁴—, —O—, —S—, —SO—, —SO₂—, —P(R⁴)—, —P(═O)(R⁴)—, —N(R⁴)SO₂—,—SO₂N(R⁴)—, —C(═S)—, —C(═NR⁴)—, —N═C— or —C(═N₂)—;

L¹ may be —(C(R^(5a))(R^(5b)))-, and n may be selected from the groupconsisting of integers ≥1;

wherein 0 or no less than 1 methylene unit of L¹ may be independentlyreplaced by -Cy-, —N(R⁶)C(O)—, —C(O)N(R⁶)—, —C(O)—, —OC(O)—, —C(O)O—,—NR⁶—, —O—, —S—, —SO—, —SO₂—, —P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—,—SO₂N(R⁶)—, —C(═S)—, —C(═NR⁶)—, —N═C— or —C(═N₂)—;

-Cy- may be selected from the group consisting of: 6-10 memberedarylene, 5-8 membered heteroarylene, 3-10 membered heterocyclylene, and3-10 membered saturated or partially unsaturated carbocyclylene, wherein-Cy- is unsubstituted or may be independently substituted with no lessthan 1 substituent R⁷;

wherein each R^(3a), each R^(3b), each R⁴, each R^(5a), each R^(5b) andeach R⁶ may each independently be hydrogen, protium, deuterium, tritium,halogen, —NO₂, —CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R,—C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group whichmay be optionally substituted with R;

wherein each R², each R⁷ and each R⁸ may each independently be hydrogen,protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R,or a C₁₋₆ aliphatic group which may be optionally substituted with R;

wherein each R, each R^(a) and each R^(b) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(I-A):

for example, wherein R^(3a) and R^(5a), R⁴ and R^(5a), R^(3a) and R⁶ orR⁴ and R⁶ may each independently optionally form a ring B together withan atom therebetween, wherein the ring B may be selected from the groupconsisting of: 5-8 membered heteroarylene and 3-10 membered saturated orpartially unsaturated heterocyclylene, and the ring B is unsubstitutedor may be substituted with no less than 1 substituent R⁸; each R^(3a),each R^(3b), each R⁴, each R^(5a), each R^(5b) and each R⁶ may eachindependently be hydrogen, protium, deuterium, tritium, halogen, —NO₂,—CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R,—C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group whichmay be optionally substituted with R; or R^(3a) and R^(5a), R⁴ andR^(5a), R^(3a) and R⁶ or R⁴ and R⁶ may each independently optionallyform a ring B together with an atom therebetween, wherein the ring B maybe selected from the group consisting of: 5-8 membered heteroarylene and3-10 membered saturated or partially unsaturated heterocyclylene, andthe ring B is unsubstituted or may be substituted with no less than 1substituent R⁸;

for example, wherein R^(3a) and R^(5a) may form a ring B together withan atom therebetween, wherein the ring B may be selected from the groupconsisting of: 5-8 membered heteroarylene and 3-10 membered saturated orpartially unsaturated heterocyclylene, and the ring B is unsubstitutedor may be substituted with no less than 1 substituent R⁸; each R^(3b),each R⁴, each R^(5b) and each R⁶ may each independently be hydrogen,protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R,or a C₁₋₆ aliphatic group which may be optionally substituted with R; orR^(3a) and R^(5a), R⁴ and R^(5a), R^(3a) and R⁶ or R⁴ and R⁶ may eachindependently optionally form a ring B together with an atomtherebetween, wherein the ring B may be selected from the groupconsisting of: 5-8 membered heteroarylene and 3-10 membered saturated orpartially unsaturated heterocyclylene, and the ring B is unsubstitutedor may be substituted with no less than 1 substituent R⁸;

for example, wherein R⁴ and R^(5a) may form a ring B together with anatom therebetween, wherein the ring B may be selected from the groupconsisting of: 5-8 membered heteroarylene and 3-10 membered saturated orpartially unsaturated heterocyclylene, and the ring B is unsubstitutedor may be substituted with no less than 1 substituent R⁸; each R^(3a),each R^(3b), each R^(5b) and each R⁶ may each independently be hydrogen,protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R,or a C₁₋₆ aliphatic group which may be optionally substituted with R; orR^(3′) and R^(5a), R⁴ and R^(5a), R^(3a) and R⁶ or R⁴ and R⁶ may eachindependently optionally form a ring B together with an atomtherebetween, wherein the ring B may be selected from the groupconsisting of: 5-8 membered heteroarylene and 3-10 membered saturated orpartially unsaturated heterocyclylene, and the ring B is unsubstitutedor may be substituted with no less than 1 substituent R⁸;

for example, wherein R^(3a) and R⁶ may form a ring B together with anatom therebetween, wherein the ring B may be selected from the groupconsisting of: 5-8 membered heteroarylene and 3-10 membered saturated orpartially unsaturated heterocyclylene, and the ring B is unsubstitutedor may be substituted with no less than 1 substituent R⁸; each R^(3b),each R⁴, each R^(5a) and each R^(5b) may each independently be hydrogen,protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R,or a C₁₋₆ aliphatic group which may be optionally substituted with R; orR^(3′) and R^(5a), R⁴ and R^(5a), R^(3a) and R⁶ or R⁴ and R⁶ may eachindependently optionally form a ring B together with an atomtherebetween, wherein the ring B may be selected from the groupconsisting of: 5-8 membered heteroarylene and 3-10 membered saturated orpartially unsaturated heterocyclylene, and the ring B is unsubstitutedor may be substituted with no less than 1 substituent for example,wherein R⁴ and R⁶ may independently optionally form a ring B togetherwith an atom therebetween, wherein the ring B may be selected from thegroup consisting of: 5-8 membered heteroarylene and 3-10 memberedsaturated or partially unsaturated heterocyclylene, and the ring B isunsubstituted or may be substituted with no less than 1 substituent R⁸;each R^(3a), each R^(3b), each R^(5a) and each R^(5b) may eachindependently be hydrogen, protium, deuterium, tritium, halogen, —NO₂,—CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R,—C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group whichmay be optionally substituted with R; or lea and R^(5a), R⁴ and R^(5a),R^(3a) and R⁶ or R⁴ and R⁶ may each independently optionally form a ringB together with an atom therebetween, wherein the ring B may be selectedfrom the group consisting of: 5-8 membered heteroarylene and 3-10membered saturated or partially unsaturated heterocyclylene, and thering B is unsubstituted or may be substituted with no less than 1substituent R⁸;

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(I-A):

wherein, R¹ may be —O—;

L² may be —(C(R^(3a))(R^(3b)))_(m)—R, and m may be selected from thegroup consisting of integers from 1 to 3;

wherein 0 methylene units of L² may be replaced;

L¹ may be —(C(R^(5a))(R^(5b)))_(n)—, and n may be selected from thegroup consisting of integers from 2 to 4;

wherein 0, 1 or 2 methylene units of L¹ may be replaced by —N(R⁶)C(O)—,—C(O)—, —OC(O)—, —NR⁶—, —O— or —C(═S)—;

wherein each R^(3a), each R^(3b), each R^(5a), each R^(5b) and each R⁶may each independently be hydrogen, halogen, or a C₁₋₆ aliphatic groupwhich may be optionally substituted with R; or R^(3a) and R^(5a) mayform a ring B together with an atom therebetween, wherein the ring B maybe selected from 5 membered saturated heterocyclylene, and the ring B isunsubstituted;

R may be hydrogen or halogen.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(I-A):

wherein, R¹ may be —O—;

L² may be —(C(R^(3a))(R^(3b)))_(m)—R, and m may be selected from thegroup consisting of integers 1 and 2;

wherein 0 methylene units of L² may be replaced;

L¹ may be —(C(R^(5a))(R^(5b)))_(n)—, and n may be selected from thegroup consisting of integers 2 and 3;

wherein 0 or 1 methylene unit of L¹ may be replaced by —C(O)—;

wherein each R^(3a), each R^(3b), each R^(5a) and each R^(5b) may eachindependently be hydrogen; or R^(3a) and R^(5a) may form a ring Btogether with an atom therebetween, wherein the ring B may be selectedfrom 5 membered saturated heterocyclylene having 1 nitrogen heteroatom,and the ring B is unsubstituted;

R may be hydrogen.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(I-A):

wherein, R¹ may be —O—;

L² may be —(C(R^(3a))(R^(3b)))_(m)—R, and m may be selected from thegroup consisting of integers 1 and 2;

wherein 0 methylene units of L² may be replaced;

L¹ may be —(C(R^(5a))(R^(5b)))₂—;

wherein 1 methylene unit of L¹ may be replaced by —C(O)—;

wherein each R^(3a), each R^(3b), each R^(5a) and each R^(5b) may eachindependently be hydrogen; or R^(3a) and R^(5a) may form a ring Btogether with an atom therebetween, wherein the ring B may be selectedfrom 5 membered saturated heterocyclylene having 1 nitrogen heteroatom,and the ring B is unsubstituted;

R may be hydrogen.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise the following group ofstructures:

wherein, R¹ may be selected from the group consisting of: —O—, —HN—,—P(═O)H— and —S—.

In another embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(II-A):

wherein, X¹ may be selected from the group consisting of: n, P, andsaturated or unsaturated C; when X¹ may be saturated C, X¹ may besubstituted with IV; ring A optionally links the structure shown asformula (II-A) to other molecular moieties;

when X¹ may be saturated C, ring A may be selected from the groupconsisting of: 3-10 membered saturated or partially unsaturatedheterocyclyl, and 3-10 membered saturated or partially unsaturatedcarbocyclyl, wherein ring A may be substituted with 0 or no less than 1substituent R^(1a);

or, when X¹ may be unsaturated C, ring A may be selected from the groupconsisting of: 6-10 membered aryl, 5-8 membered heteroaryl, 3-10membered partially unsaturated heterocyclyl, and 3-10 membered partiallyunsaturated carbocyclyl, wherein ring A may be substituted with 0 or noless than 1 substituent R^(1b);

or, when X¹ may be N or P, ring A may be selected from the groupconsisting of: 5-8 membered heteroaryl and 3-10 membered saturated orpartially unsaturated heterocyclyl, wherein ring A may be substitutedwith 0 or no less than 1 substituent R^(1c);

when ring A may be selected from the group consisting of: 6-10 memberedaryl, 5-8 membered heteroaryl, and 3-10 membered saturated or partiallyunsaturated carbocyclyl, ring A may be substituted with p L², wherein L²cannot be R^(n);

or, when ring A may be 3-10 membered saturated or partially unsaturatedheterocyclyl, ring A may be substituted with p L², or ring A maycomprise q ring-forming heteroatom X², and X² is used to link thestructure shown as formula (II-A) to other molecular moieties;

X² may be selected from the group consisting of: N and P;

L² may be —R²-L³-, and R² is used to link the structure shown as formula(II-A) to other molecular moieties;

L³ may be —(C(R^(3a))(R^(3b)))_(m)—, wherein when L³ may comprise amethylene unit, 0 or no less than 1 methylene unit of L³ may beindependently replaced by —N(R⁴)C(O)—, —C(O)N(R⁴)—, —C(O)—, —OC(O)—,—C(O)O—, —NR⁴—, —O—, —S—, —SO—, —SO₂—, —P(R⁴)—, —P(═O)(R⁴)—, —N(R⁴)SO₂—,—SO₂N(R⁴)—, —C(═S)—, —C(═NR⁴)—, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

R² may be selected from the group consisting of: —O—, —(R^(2a))N—, —S—and —P(═O)(R^(2a))—;

L¹ may be —(C(R^(5a))(R^(5b)))_(n)—, wherein when L¹ may comprise amethylene unit, 0 or no less than 1 methylene unit of L¹ may beindependently replaced by —N(R⁶)C(O)—, —C(O)N(R⁶)—, —C(O)—, —OC(O)—,—C(O)O—, —NR⁶—, —O—, —S—, —SO—, —SO₂—, —P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—,—SO₂N(R⁶)—, —C(═S)—, —C(═NR⁶)—, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

wherein each R^(1a), each R^(1b), each R^(1c), each R^(2a), each R^(3a),each R^(3b), each R⁴, each R^(5a), each R^(5b) and each R⁶ may eachindependently be hydrogen, protium, deuterium, tritium, halogen, —NO₂,—CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R,—C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group whichmay be optionally substituted with R;

wherein each R, each R^(a) and each R^(b) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

m and n may each independently be selected from the group consisting ofintegers ≥0, and p and q may each independently be selected from thegroup consisting of integers ≥1.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(II-Ax):

wherein, X¹ may be saturated C, and X¹ may be substituted with R^(n);

ring A may be selected from the group consisting of: 3-10 memberedsaturated or partially unsaturated heterocyclyl and 3-10 memberedsaturated or partially unsaturated carbocyclyl, wherein ring A may besubstituted with 0 or no less than 1 substituent R^(1a);

ring A may be substituted with p L², wherein p may be selected from thegroup consisting of integers ≥1, and L² can not be R^(n);

L² may be —R²-L³-;

L³ may be —(C(R^(3a))(R^(3b)))_(m), and m may be selected from the groupconsisting of integers ≥0;

wherein when L³ may comprise a methylene unit, 0 or no less than 1methylene unit of L³ may be independently replaced by —N(R⁴)C(O)—,—C(O)N(R⁴)—, —C(O)—, —OC(O)—, —C(O)O—, —NR⁴—, —O—, —S—, —SO—, —SO₂—,—P(R⁴)—, —P(═O)(R⁴)—, —N(R⁴)SO₂—, —SO₂N(R⁴)—, —C(═S)—, —C(═NR⁴)—, —N═N—,—C═N—, —N═C— or —C(═N₂)—;

R² may be selected from the group consisting of: —O—, —(R^(2′))N—, —S—and —P(═O)(R^(2a))—;

L¹ may be —(C(R^(5a))(R^(5b)))_(n)—, and n may be selected from thegroup consisting of integers ≥0;

wherein when L¹ may comprise a methylene unit, 0 or no less than 1methylene unit of 12 may be independently replaced by —N(R⁶)C(O)—,—C(O)N(R⁶)—, —C(O)—, —OC(O)—, —C(O)O—, —NR⁶—, —O—, —S—, —SO—, —SO₂—,—P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—, —SO₂N(R⁶)—, —C(═S)—, —C(═NR⁶)—, —N═N—,—C═N—, —N═C— or —C(═N₂)—;

wherein each R^(1a), each R^(2a), each R^(3a), each R^(3b), each R⁴,each R^(5a), each R^(5b), each R⁶ and each IV may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R,or a C₁₋₆ aliphatic group which may be optionally substituted with R;

wherein each R, each R^(a) and each R^(b) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(II-Ay):

wherein, X¹ may be saturated C, and X¹ may be substituted with IV;

ring A may be 3-10 membered saturated or partially unsaturatedheterocyclyl, and ring A can not be substituted or may be substitutedwith no less than 1 substituent R^(1a);

ring A may comprise q ring-forming heteroatom X², and X² is used fordirect or indirect linking of a ligand; q may be selected from the groupconsisting of integers ≥1, and X² may be selected from the groupconsisting of: N and P;

L¹ may be —(C(R^(5a))(R^(5b)))_(n)—, and n may be selected from thegroup consisting of integers ≥0;

wherein 0 or no less than 1 methylene unit of L¹ may be independentlyreplaced by —N(R⁶)C(O)—, —C(O)N(R⁶)—, —C(O)—, —OC(O)—, —C(O)O—, —NR⁶—,—O—, —S—, —SO—, —SO₂—, —P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—, —SO₂N(R⁶)—,—C(═S)—, —C(═NR⁶)—, —N═C— or —C(═N₂)—;

wherein each R^(1a), each R^(5a), each R^(5b), each R⁶ and each R^(n)may each independently be hydrogen, protium, deuterium, tritium,halogen, —NO₂, —CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R,—C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group whichmay be optionally substituted with R;

wherein each R, each R^(a) and each R^(b) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(II-Ax):

wherein, X¹ may be unsaturated C;

ring A may be selected from the group consisting of: 6-10 membered aryl,5-8 membered heteroaryl, 3-10 membered partially unsaturatedheterocyclyl, and 3-10 membered partially unsaturated carbocyclyl, andring A can not be substituted or may be substituted with no less than 1substituent R^(1b);

ring A may be substituted with p L², wherein p may be selected from thegroup consisting of integers ≥1;

L² may be —R²-L³-, and R² is used for direct or indirect linking of aligand;

L³ may be —(C(R^(3a))(R^(3b)))_(m), and m may be selected from the groupconsisting of integers ≥0;

wherein 0 or no less than 1 methylene unit of L³ may be independentlyreplaced by —N(R⁴)C(O)—, —C(O)N(R⁴)—, —C(O)—, —OC(O)—, —C(O)O—, —O—,—S—, —SO—, —SO₂—, —P(R⁴)—, —P(═O)(R⁴)—, —N(R⁴)SO₂—, —SO₂N(R⁴)—, —C(═S)—,—C(═NR⁴)—, —N═C— or —C(═N₂)—;

R² may be selected from the group consisting of: —O—, —(R^(2a))N—, —S—and —P(═O)(R^(2a))—;

L¹ may be —(C(R^(5a))(R^(5b)))_(n)—, and n may be selected from thegroup consisting of integers ≥0;

wherein 0 or no less than 1 methylene unit of L¹ may be independentlyreplaced by —N(R⁶)C(O)—, —C(O)N(R⁶)—, —C(O)—, —OC(O)—, —C(O)O—, —NR⁶—,—O—, —S—, —SO—, —SO₂—, —P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—, —SO₂N(R⁶)—,—C(═S)—, —C(═NR⁶)—, —N═C— or —C(═N₂)—;

wherein each R^(1b), each R^(2a), each R^(3a), each R^(3b), each R⁴,each R^(5a), each R^(5b) and each R⁶ may each independently be hydrogen,protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R,or a C₁₋₆ aliphatic group which may be optionally substituted with R;

wherein each R, each R^(a) and each R^(b) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(II-Ay):

wherein, X¹ may be unsaturated C;

ring A may be 3-10 membered partially unsaturated heterocyclyl, and ringA can not be substituted or may be substituted with no less than 1substituent R^(1b);

ring A may comprise q ring-forming heteroatom X², and X² is used fordirect or indirect linking of a ligand; q may be selected from the groupconsisting of integers ≥1, and X² may be selected from the groupconsisting of: N and P;

L¹ may be —(C(R^(5a))(R^(5b)))_(n)—, and n may be selected from thegroup consisting of integers ≥0;

wherein 0 or no less than 1 methylene unit of L¹ may be independentlyreplaced by —N(R⁶)C(O)—, —C(O)N(R⁶)—, —C(O)—, —OC(O)—, —C(O)O—, —NR⁶—,—O—, —S—, —SO—, —SO₂—, —P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—, —SO₂N(R⁶)—,—C(═S)—, —C(═NR⁶)—, —N═C— or —C(═N₂)—;

wherein each R^(1b), each R^(5a), each R^(5b) and each R⁶ may eachindependently be hydrogen, protium, deuterium, tritium, halogen, —NO₂,—CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R,—C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group whichmay be optionally substituted with R;

wherein each R, each R^(a) and each R^(b) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(II-Ax):

wherein, X¹ may be N or P;

ring A may be selected from the group consisting of: 5-8 memberedheteroaryl and 3-10 membered saturated or partially unsaturatedheterocyclyl, and ring A can not be substituted or may be substitutedwith no less than 1 substituent R^(1c);

ring A may be substituted with p L², wherein p may be selected from thegroup consisting of integers ≥1;

L² may be —R²-L³-, and R² is used for direct or indirect linking of aligand;

L³ may be —(C(R^(3a))(R^(3b)))_(m), and m may be selected from the groupconsisting of integers ≥0;

wherein 0 or no less than 1 methylene unit of L³ may be independentlyreplaced by —N(R⁴)C(O)—, —C(O)N(R⁴)—, —C(O)—, —OC(O)—, —C(O)O—, —O—,—S—, —SO—, —SO₂—, —P(R⁴)—, —P(═O)(R⁴)—, —N(R⁴)SO₂—, —SO₂N(R⁴)—, —C(═S)—,—C(═NR⁴)—, —N═C— or —C(═N₂)—;

R² may be selected from the group consisting of: —O—, —(R^(2a))N—, —S—and —P(═O)(R^(2a))—;

L¹ may be —(C(R^(5a))(R^(5b)))_(n)—, and n may be selected from thegroup consisting of integers ≥0;

wherein 0 or no less than 1 methylene unit of L¹ may be independentlyreplaced by —N(R⁶)C(O)—, —C(O)N(R⁶)—, —C(O)—, —OC(O)—, —C(O)O—, —O—,—S—, —SO—, —SO₂—, —P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—, —SO₂N(R⁶)—, —C(═S)—,—C(═NR⁶)—, —N═C— or —C(═N₂)—;

wherein each R^(1c), each R^(2a), each R^(3a), each R^(3b), each R⁴,each R^(5a), each R^(5b) and each R⁶ may each independently be hydrogen,protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R,or a C₁₋₆ aliphatic group which may be optionally substituted with R;

wherein each R, each R^(a) and each R^(b) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(II-Ay):

wherein, X¹ may be N or P;

ring A may be 3-10 membered saturated or partially unsaturatedheterocyclyl, and ring A can not be substituted or may be substitutedwith no less than 1 substituent R^(1c);

ring A may comprise q ring-forming heteroatom X², and X² is used fordirect or indirect linking of a ligand; q may be selected from the groupconsisting of integers ≥1, and X² may be selected from the groupconsisting of: N and P;

L¹ may be —(C(R^(5a))(R^(5b)))_(n)—, and n may be selected from thegroup consisting of integers ≥0;

wherein 0 or no less than 1 methylene unit of L¹ may be independentlyreplaced by —N(R⁶)C(O)—, —C(O)N(R⁶)—, —C(O)—, —OC(O)—, —C(O)O—, —NR⁶—,—O—, —S—, —SO—, —SO₂—, —P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—, —SO₂N(R⁶)—,—C(═S)—, —C(═NR⁶)—, —N═C— or —C(═N₂)—;

wherein each R^(1c), each R^(5a), each R^(5b) and each R⁶ may eachindependently be hydrogen, protium, deuterium, tritium, halogen, —NO₂,—CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R,—C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group whichmay be optionally substituted with R;

wherein each R, each R^(a) and each R^(b) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(II-Ax):

wherein, X¹ may be saturated C, and X¹ may be substituted with IV;

ring A may be selected from the group consisting of: 3-6 memberedsaturated heterocyclyl and 3-6 membered saturated or partiallyunsaturated carbocyclyl;

p may be 1, and L² can not be R^(n);

L² may be —R²-L³-;

L³ may be —(C(R^(3a))(R^(3b)))_(m)—, and m may be selected from thegroup consisting of integers from 0 to 2, wherein when L³ may comprise amethylene unit, 0 or 1 methylene unit of L³ may be replaced by —C(O)— or—C(═S)—;

R² may be selected from —O—;

L¹ may be —(C(R^(5a))(R^(5b)))_(n)—, and n may be selected from thegroup consisting of 0 and 1;

wherein when L¹ may comprise a methylene unit, 0 or 1 methylene unit ofL¹ may be replaced by —C(O)— or —C(═S)—;

wherein each R^(3a), each R^(3b), each R^(5a), each R^(5b) and eachR^(n) may each independently be hydrogen, halogen, or a C₁₋₆ aliphaticgroup which may be optionally substituted with R;

wherein each R may independently be hydrogen or halogen.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(II-Ax):

wherein, X¹ may be saturated C, and X¹ is linked to R^(n), wherein R^(n)may be H;

ring A may be selected from the group consisting of: 5 memberedsaturated heterocyclyl having 1 nitrogen heteroatom, and 4-6 memberedsaturated carbocyclyl;

P may be 1;

L² may be —R²-L³-, and L³ is directly linked to ring A;

L³ may be —(C(R^(3a))(R^(3b)))_(m)—, and m may be 0 or 2;

R² may be —O—;

L¹ may be —C(O)—;

wherein each R^(3a) and each R^(3b) may each independently be hydrogen.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(II-Ay):

wherein, X¹ may be saturated C, and X¹ may be substituted with R^(n);

ring A may be 3-6 membered saturated heterocyclyl;

ring A may comprise 1 ring-forming heteroatom N, and N is used fordirect or indirect linking of a ligand;

L¹ may be —(C(R^(5a))(R^(5b)))_(n)—, and n may be selected from thegroup consisting of 0 and 1;

wherein 0 or 1 methylene unit of L¹ may be replaced by —C(O)— or—C(═S)—;

wherein each R^(5a), each R^(5b) and each IV may each independently behydrogen, halogen, or a C₁₋₆ aliphatic group which may be optionallysubstituted with R;

wherein each R may independently be hydrogen or halogen.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(II-Ay):

wherein, X¹ may be saturated C, and X¹ may be substituted with H;

ring A may be 5 membered saturated heterocyclyl having 1 heteroatom N;

ring A may comprise 1 ring-forming heteroatom N, and N is used fordirect or indirect linking of a ligand;

L¹ may be —C(O)—.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(II-Ax):

wherein, X¹ may be unsaturated C;

ring A may be selected from the group consisting of: 6 membered aryl,5-8 membered heteroaryl, 3-10 membered partially unsaturatedheterocyclyl, and 3-10 membered partially unsaturated carbocyclyl, andring A can not be substituted or may be independently substituted with 1substituent R^(1b);

P may be 1;

L² may be —R²-L³-, and R² is used for direct or indirect linking of aligand;

L³ may be —C(R^(3a))(R^(3b))—;

R² may be selected from the group consisting of: —O—, —(R^(2a))N- and—S—;

L¹ may be —C(R^(5a))(R^(5b))—;

wherein 0 or 1 methylene unit of L¹ may be replaced by —C(O)— or—C(═S)—;

wherein each R^(1b), each R^(2a), each R^(3a), each R^(3b), each R^(5a)and each R^(5b) may each independently be hydrogen, halogen, or a C₁₋₆aliphatic group which may be optionally substituted with R;

wherein each R may independently be hydrogen or halogen.

For example, the compound may comprise a structure shown as formula(II-Ax):

wherein, X¹ may be unsaturated C;

ring A may be selected from the group consisting of: 6 membered aryl and5-8 membered heteroaryl;

P may be 1;

L² may be —R²-L³-, and R² is used for direct or indirect linking of aligand;

L³ may be —C(R^(3a))(R^(3b))—;

R² may be selected from the group consisting of: —O—, —(R^(2a))N- and—S—;

L¹ may be —C(R^(5a))(R^(5b))—;

wherein 0 or 1 methylene unit of L¹ may be replaced by —C(O)— or—C(═S)—;

wherein each R^(2a), each R^(3a), each R^(3b), each R^(5a) and eachR^(5b) may each independently be hydrogen, halogen, or a C₁₋₆ aliphaticgroup which may be optionally substituted with R;

wherein each R may independently be hydrogen or halogen.

For example, the compound may comprise a structure shown as formula(II-Ax):

wherein, X¹ may be unsaturated C;

ring A may be 6 membered aryl;

P may be 1;

L² may be —R²-L³-, and R² is used for direct or indirect linking of aligand;

L³ may be —C(R^(3a))(R^(3b))—;

R² may be —O—;

L¹ may be —C(O)—;

wherein each R^(3a), each R^(3b), each R^(5a) and each R^(5b) may eachindependently be hydrogen or a C₁₋₆ aliphatic group;

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(II-Ay):

wherein, X¹ may be unsaturated C;

ring A may be 5 membered partially unsaturated heterocyclyl;

ring A may comprise 1 ring-forming heteroatom N, and N is used fordirect or indirect linking of a ligand;

L¹ may be —C(R^(5a))(R^(5b)),

wherein 0 or 1 methylene unit of 12 may be replaced by —C(O)— or—C(═S)—;

wherein each R^(5a) and each R^(5b) may each independently be hydrogen,halogen, or a C₁₋₆ aliphatic group which may be optionally substitutedwith R;

wherein each R may independently be hydrogen or halogen.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(II-Ax):

wherein, X¹ may be N;

ring A may be 6 membered saturated heterocyclyl;

P may be 1;

L² may be —R²-L³-, and R² is used for direct or indirect linking of aligand;

L³ may be —(C(R^(3a))(R^(3b)))_(m)—, and m may be 1 or 2,

wherein 0 or 1 methylene unit of L³ may be replaced by —C(O)— or—C(═S)—;

R² may be selected from the group consisting of: —O—, —(R^(2a))N- and—S—;

L¹ may be —C(R^(5a))(R^(5b))—,

wherein 1 methylene unit of L¹ may be replaced by —C(O)— or —C(═S)—;

wherein each R^(2a), each R^(3a), each R^(3b), each R^(5a) and eachR^(5b) may each independently be hydrogen, halogen, or a C₁₋₆ aliphaticgroup which may be optionally substituted with R;

wherein each R may independently be hydrogen or halogen.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(II-Ay):

wherein, X¹ may be N;

ring A may be 5 membered partially unsaturated heterocyclyl;

ring A may comprise 1 ring-forming heteroatom N, and N is used fordirect or indirect linking of a ligand;

L¹ may be —C(R^(5a))(R^(5b)),

wherein no less than 1 methylene unit of L¹ may be replaced by —C(O)— or—C(═S)—;

wherein each R^(5a) and each R^(5b) may each independently be hydrogen,halogen, or a C₁₋₆ aliphatic group which may be optionally substitutedwith R;

wherein each R may independently be hydrogen or halogen.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise the following group ofstructures:

wherein, R² may be selected from the group consisting of: —O—, —HN—,—P(═O)H— and —S—.

In another embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be selected from the group consisting of: —O—, —(R²)N—,—P(═O)(R²)— and —S—;

X may be selected from the group consisting of:-L¹-C(R^(1a))(R^(1b))—C(O)—, —L¹-C(R^(1a))(R^(1b))—C(S)—, -L¹-L⁰- and-L³-L²-;

L¹ may be —(C(R^(3a))(R^(3b)))_(m)—, wherein when L¹ may comprise amethylene unit, 0 or no less than 1 methylene unit of L¹ may beindependently replaced by —C(O)— —C(═S)—, —C(═NR^(4b))— or —C(═N₂)—;

L⁰ may be —C(R^(2a))(R^(2b))—, or L⁰ may be —C(═S)—, —C(═NR^(4a))— or—C(═N₂)—;

L² may be —C(R^(5a))(R^(5b))—, wherein 0 or 1 methylene unit of L² maybe replaced by —N(R⁶)C(O)—, —C(O)N(R⁶)—, —C(O)—, —OC(O)—, —C(O)O—,—NR⁶—, —O—, —S—, —SO—, —SO₂—, —P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—,—SO₂N(R⁶)—, —C(═S)—, —C(═NR⁶)—, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

L³ may be —(C(R^(7a))(R^(7b)))_(n)—, wherein no less than 1 methyleneunit of L³ may be independently replaced by —N(R⁸)C(O)—, —C(O)N(R⁸)—,—OC(O)—, —C(O)O—, —NR⁸—, —O—, —S—, —SO—, —SO₂—, —P(R⁸)—, —P(═O)(R⁸)—,—N(R⁸)SO₂—, —SO₂N(R⁸)—, —N═N—, —C═N— or —N═C—, and 0 or no less than 1methylene unit of L³ may also independently be replaced by —C(O)—,—C(═S)—, —C(═NR⁸)— or —C(═N₂)—;

wherein each R^(1a), each R^(1b), each R², each R^(2a), each R^(2b),each R^(3a), each R^(3b), each R^(4a), each R^(4b), each R^(5a), eachR^(5b), each R⁶, each R^(7a), each R^(7b) and each R⁸ may eachindependently be hydrogen, protium, deuterium, tritium, halogen, —NO₂,—CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R,—C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group whichmay be optionally substituted with R;

wherein each R, each R^(a) and each R^(b) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

m may be selected from the group consisting of integers ≥0, and n may beselected from the group consisting of integers ≥1;

when R¹ may be —O— or —HN—, and X may be -L¹-CH₂—C(O)—, and when L¹ maycomprise a methylene unit, no less than 1 methylene unit of L¹ may beindependently replaced by —C(O)—, —C(═S)—, —C(═NR^(4b))— or —C(═N₂)—, orR^(3a) and R^(3b) can not be both hydrogen in each —C(R^(3a))(R^(3b))-of L¹;

when R¹ may be —HN—, X may be -L¹-L⁰-, and L⁰ may be —CH₂—, no less than1 methylene unit of L¹ may be independently replaced by —C(O)—, —C(═S)—,—C(═NR^(4b))— or —C(═N₂)—, or each R^(3a) and each R^(3b) can not beboth hydrogen;

when R¹ may be —O—, X may be -L³-C(O)—, and 1 methylene unit of L³ maybe replaced by —NR⁸, R⁸ can not be —CH₂—CH₂—NH₂;

when R¹ may be —NH—, and X may be -L³-C(O)—, no less than 1 methyleneunit of L³ may be replaced by —N(R⁸)C(O)—, —OC(O)—, —C(O)O—, —S—, —SO—,—SO₂—, —P(R⁸)—, —P(═O)(R⁸)—, —N(R⁸)SO₂—, —SO₂N(R⁸)—, —N═N—, —C═N— or—N═C—.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be selected from the group consisting of: —O—, —(R²)N-and —S—;

X may be -L¹-C(R^(1a))(R^(1b))—C(S)—;

L¹ may be —(C(R^(3a))(R^(3b)))_(m), and m may be selected from the groupconsisting of integers ≥0;

wherein 0 or no less than 1 methylene unit of L¹ may be independentlyreplaced by —C(O)—, —C(═S)—, —C(═NR^(4b))— or —C(═N₂)—;

R² may be halogen, —NO₂, —CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R,—C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group whichmay be optionally substituted with R;

wherein each R^(1a), each R^(1b), each R^(3a), each R^(3b) and eachR^(4b) may each independently be hydrogen, protium, deuterium, tritium,halogen, —NO₂, —CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R,—C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group whichmay be optionally substituted with R;

wherein each R, each R^(a) and each R^(b) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —S— or —(R²)N—;

X may be -L¹-C(R^(1a))(R^(1b))—C(O)—;

L¹ may be —(C(R^(3a))(R^(3b)))_(m), and m may be selected from the groupconsisting of integers ≥0;

wherein 0 or no less than 1 methylene unit of L¹ may be independentlyreplaced by —C(O)—, —C(═S)—, —C(═NR^(4b))— or —C(═N₂)—;

R² may be halogen, —NO₂, —CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R,—C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group whichmay be optionally substituted with R;

wherein each R^(1a), each R^(1b), each R^(3a), each R^(3b) and eachR^(4b) may each independently be hydrogen, protium, deuterium, tritium,halogen, —NO₂, —CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R,—C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group whichmay be optionally substituted with R;

wherein each R, each R^(a) and each R^(b) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —O— or —HN—;

X may be -L¹-CH₂—C(O)—;

L¹ may be —(C(R^(3a))(R^(3b)))^(m), and m may be selected from the groupconsisting of integers ≥1;

wherein no less than 1 methylene unit of L¹ may be independentlyreplaced by —C(O)—, —C(═S)—, —C(═NR^(4b))— or —C(═N₂)—;

wherein each R^(1a), each R^(3b) and each R^(4b) may each independentlybe hydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂Ror a C₁₋₆ aliphatic group which may be optionally substituted with R;

wherein each R, each R^(a) and each R^(b) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —O— or —HN—;

X may be -L¹-CH₂—C(O)—;

L¹ may be —(C(R^(3a))(R^(3b)))^(m), and m may be selected from the groupconsisting of integers ≥0;

R^(3a) and R^(3b) can not be both hydrogen in each —C(R^(3a))(R^(3b))—,or no less than 1 methylene unit of L¹ may be independently replaced by—C(O)— —C(═S)—, —C(═NR^(4b))— or —C(═N₂)—;

wherein each R^(3a), each R^(3b) and each R^(4b) may each independentlybe hydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂Ror a C₁₋₆ aliphatic group which may be optionally substituted with R;

wherein each R, each R^(a) and each R^(b) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —O—, —S— or —(R²)N—;

X may be -L¹-L⁰-;

L⁰ may be —C(R^(2a))(R^(2b))—, or L⁰ may be —C(═S)—, —C(═NR^(4a))— or—C(═N₂)—;

L¹ may be —(C(R^(3a))(R^(3b)))_(m), and m may be selected from the groupconsisting of integers ≥0;

wherein 0 or no less than 1 methylene unit of L¹ may be independentlyreplaced by —C(O)—, —C(═S)—, —C(═NR^(4b))— or —C(═N₂)—;

R² may be halogen, —NO₂, —CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R,—C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group whichmay be optionally substituted with R;

wherein each R^(2a), each R^(2b), R^(3a), each R^(3b), each R^(4a) andeach R^(4b) may each independently be hydrogen, protium, deuterium,tritium, halogen, —NO₂, —CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R,—C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group whichmay be optionally substituted with R;

wherein each R, each R^(a) and each R^(b) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —HN—;

X may be -L¹-L⁰-;

L⁰ may be —C(R^(2a))(R^(2b))—, or L⁰ may be —C(═S)—, —C(═NR^(4a))— or—C(═N₂)—;

L¹ may be —(C(R^(3a))(R^(3b)))_(m)—, m may be selected from the groupconsisting of integers ≥0, and each R^(3a) and each R^(3b) can not beboth hydrogen;

wherein 0 or no less than 1 methylene unit of L¹ may be independentlyreplaced by —C(O)—, —C(═S)—, —C(═NR^(4b))— or —C(═N₂)—;

wherein each R^(2a), each R^(2b), R^(3a), each R^(3b), each R^(4a) andeach R^(4b) may each independently be hydrogen, protium, deuterium,tritium, halogen, —NO₂, —CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R,—C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group whichmay be optionally substituted with R;

wherein each R, each R^(a) and each R^(b) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, X may be -L³-L²-;

L² may be —C(R^(5a))(R^(5b))—, wherein 0 or 1 methylene unit of L² maybe replaced by —N(R⁶)C(O)—, —C(O)N(R⁶)—, —C(O)—, —OC(O)—, —C(O)O—,—NR⁶—, —O—, —S—, —SO—, —SO₂—, —P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—,—SO₂N(R⁶)—, —C(═S)—, —C(═NR⁶)—, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

R¹ may be —S— or —(R²)N—; or R¹ may be —O— and L² can not be —C(O)—; orR¹ may be —NH— and L² can not be —C(O)—;

L³ may be —(C(R^(7a))(R^(7b)))_(n)—, and n may be selected from thegroup consisting of integers ≥1;

wherein no less than 1 methylene unit of L³ may be independentlyreplaced by —N(R⁸)C(O)—, —C(O)N(R⁸)—, —OC(O)—, —C(O)O—, —NR⁸—, —O—, —S—,—SO—, —SO₂—, —P(R⁸)—, —P(═O)(R⁸)—, —N(R⁸)SO₂—, —SO₂N(R⁸)—, —N═N—, —C═N—or —N═C—, and 0 or no less than 1 methylene unit of L³ may also beindependently replaced by —C(O)—, —C(═S)—, —C(═NR⁸)— or —C(═N₂)—;

R² may be halogen, —NO₂, —CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R,—C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group whichmay be optionally substituted with R;

wherein each R^(5a), each R^(5b), each R⁶, each R^(7a), each R^(7b) andeach R⁸ may each independently be hydrogen, protium, deuterium, tritium,halogen, —NO₂, —CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R,—C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group whichmay be optionally substituted with R;

wherein each R, each R^(a) and each R^(b) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —O—;

X may be -L³-L²-;

wherein L² may be —C(O)—;

L³ may be —(C(R^(7a))(R^(7b)))_(n)—, and n may be selected from thegroup consisting of integers ≥0;

wherein no less than 1 methylene unit of L³ may be independentlyreplaced by —N(R⁸)C(O)—, —C(O)N(R⁸)—, —OC(O)—, —C(O)O—, —NR⁸—, —O—, —S—,—SO—, —SO₂—, —P(R⁸)—, —P(═O)(R⁸)—, —N(R⁸)SO₂—, —SO₂N(R⁸)—, —N═N—, —C═N—or —N═C—, and 0 or no less than 1 methylene unit of L³ may also beindependently replaced by —C(O)—, —C(═S)—, —C(═NR⁸)— or —C(═N₂)—;

wherein each R^(7a), each R^(7b) and each R⁸ may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂Ror a C₁₋₆ aliphatic group which may be optionally substituted with R;

wherein R, R^(a) and R^(b) may each independently be hydrogen, protium,deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH, —NH₂, —C(O)H, —CO₂H,—C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H, —C(O)NH₂, —SO₂NH₂, —OC(O)H,—N(H)SO₂H or a C₁₋₆ aliphatic group;

when 1 methylene unit of L³ may be replaced by —NR⁸, R⁸ can not be aC₁₋₆ aliphatic group which may be substituted with —NH₂.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —HN—;

X may be -L³-L²-;

wherein L² may be —C(O)—;

L³ may be —(C(R^(7a))(R^(7b)))_(n)—, and n may be selected from thegroup consisting of integers ≥1;

no less than 1 methylene unit of L³ may be replaced by —N(R⁸)C(O)—,—OC(O)—, —C(O)O—, —S—, —SO—, —SO₂—, —P(R⁸)—, —P(═O)(R⁸)—, —N(R⁸)SO₂—,—SO₂N(R⁸)—, —N═N—, —C═N— or —N═C—, and 0 or no less than 1 methyleneunit of L³ may also be independently replaced by —C(O)N(R⁸)—, —NR⁸— or—O—, and 0 or no less than 1 methylene unit of L³ may also beindependently replaced by —C(O)—, —C(═S)—, —C(═NR⁸)— or —C(═N₂)—;

wherein each R^(7a), each R^(7b) and each R⁸ may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂Ror a C₁₋₆ aliphatic group which may be optionally substituted with R;

wherein each R, each R^(a) and each R^(b) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —O—, —S— or —(R²)N—;

X may be —L¹-C(R^(1a))(R^(1b))—C(S)—;

L¹ may be —(C(R^(3a))(R^(3b)))_(m)—, and m may be 0, 1 or 2;

wherein 0 or 1 methylene unit of L¹ may be replaced by —C(O)—;

wherein each R^(1a), each R^(1b), each R², each R^(3a) and each R^(3b)may each independently be hydrogen, or a C₁₋₆ aliphatic group which maybe optionally substituted with R;

wherein each R may be hydrogen.

For example, the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —O—;

X may be —L¹-C(R^(1a))(R^(1b))—C(S)—;

L¹ may be —(CH₂)_(m)—, and m may be 1 or 2;

wherein 0 or 1 methylene unit of L¹ may be replaced by —C(O)—;

wherein each R^(1a) and each R^(1b) may each independently be hydrogen,or a C₁₋₆ aliphatic group which may be optionally substituted with R;

wherein each R may be hydrogen.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —S— or —(R²)N—;

X may be —L¹-C(R^(1a))(R^(1b))—C(O)—;

L¹ may be —(C(R^(3a))(R^(3b)))_(m)—, and m may be 0, 1 or 2;

wherein 0 or 1 methylene unit of L¹ may be replaced by —C(O)—;

R² may be a C₁₋₆ aliphatic group;

wherein each R^(1a), each R^(1b), each R^(3a) and each R^(3b) may eachindependently be hydrogen or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —S— or —(R²)N—;

X may be -L¹-C(R^(1a))(R^(1b))—C(O)—;

L¹ may be —(C(R^(3a))(R^(3b)))_(m)—, and m may be 1 or 2;

wherein 0 or 1 methylene unit of 12 may be replaced by —C(O)—;

R² may be a C₁₋₆ aliphatic group;

wherein each R^(1a), each R^(1b), each R^(3a) and each R^(3b) may eachindependently be hydrogen or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, when R¹ may be —S— or —(R²)N—,

X may be —L¹-C(R^(1a))(R^(1b))—C(O)—,

L¹ may be —(C(R^(3a))(R^(3b)))_(m)—, and m may be 0, 1 or 2,

0 or 1 methylene unit of L¹ may be replaced by —C(O)—;

R² may be a C₁₋₆ aliphatic group;

wherein each R^(1a), each R^(1b), each R^(3a) and each R^(3b) may eachindependently be hydrogen or a C₁₋₆ aliphatic group;

or, when R¹ may be —O—,

X may be -L¹-CH₂—C(O)—, and

L¹ may be —(C(R^(3a))(R^(3b)))₂—,

0 or 1 methylene unit of L¹ may be replaced by —C(O)—;

wherein each R^(3a) and each R^(3b) may each independently be hydrogenor a C₁₋₆ aliphatic group;

each R^(3a) and each R^(3b) can not be both hydrogen, or 1 methyleneunit of L¹ may be replaced by —C(O)—;

wherein each R^(3a) and each R^(3b) may each independently be hydrogenor a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —O—;

X may be -L¹-CH₂—C(O)—;

L¹ may be —(C(R^(3a))(R^(3b)))₂—;

wherein 0 or 1 methylene unit of 12 may be replaced by —C(O)—;

wherein each R^(3a) and each R^(3b) may each independently be hydrogenor a C₁₋₆ aliphatic group;

each R^(3a) and each R^(3b) can not be both hydrogen, or 1 methyleneunit of 12 may be replaced by —C(O)—;

wherein each R^(3a) and each R^(3b) may each independently be hydrogenor a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, when R¹ may be —S— or —(R²)N—,

X may be —Li—C(R^(1a))(R^(1b))—C(O)—,

L¹ may be —(C(R^(3a))(R^(3b)))_(m)—, and m may be 0, 1 or 2,

0 or 1 methylene unit of L¹ may be replaced by —C(O)—;

R² may be a C₁₋₆ aliphatic group;

wherein each R^(1a), each R^(1b), each R^(3a) and each R^(3b) may eachindependently be hydrogen or a C₁₋₆ aliphatic group;

or, when R¹ may be —O—, and

X may be -L¹-CH₂—C(O)—,

L¹ may be —C(R^(3a))(R^(3b))—, and R^(3a) and R^(3b) can not be bothhydrogen in each —C(R^(3a))(R^(3b))—;

wherein each R^(3a) and each R^(3b) may each independently be hydrogenor a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —O—;

X may be -L¹-CH₂—C(O)—;

L¹ may be —(C(R^(3a))(R^(3b)))₂—;

wherein 0 or 1 methylene unit of L¹ may be replaced by —C(O)—;

wherein each R^(3a) and each R^(3b) may each independently be hydrogenor a C₁₋₆ aliphatic group;

R^(3a) and R^(3b) can not be both hydrogen in each —C(R^(3a))(R^(3b)),or 1 methylene unit of L¹ may be replaced by —C(O)—;

wherein each R^(3a) and each R^(3b) may each independently be hydrogenor a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —O—;

X may be -L¹-CH₂—C(O)—;

L¹ may be —(CH₂)₂—;

wherein 1 methylene unit of L¹ may be replaced by —C(O)—.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —O—;

X may be -L¹-CH₂—C(O)—;

L¹ may be —C(R^(3a))(R^(3b))—_(m)-, m may be selected from the groupconsisting of integers from 1 to 5, and R^(3a) and R^(3b) can not beboth hydrogen in each —C(R^(3a))(R^(3b))—;

wherein each R^(3a) and each R^(3b) may each independently be hydrogen,halogen, or a C₁₋₆ aliphatic group which may be optionally substitutedwith R;

wherein each R may be hydrogen or halogen.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —O—;

X may be -L¹-CH₂—C(O)—;

L¹ may be —C(R^(3a))(R^(3b))—, and R^(3a) and R^(3b) can not be bothhydrogen;

wherein each R^(3a) and each R^(3b) may each independently be hydrogenor a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —O— or —(R²)N—;

X may be -L¹-L⁰-;

L⁰ may be —CH₂—, or L⁰ may be —C(═S)—;

L¹ may be —(CH₂)_(m)—, and m may be selected from the group consistingof integers from 0 to 2;

wherein 0 or 1 methylene unit of L¹ may be replaced by —C(O)— or—C(═S)—;

R² may be a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —NH—;

X may be -L¹-L⁰-;

L⁰ may be —CH₂—, or L⁰ may be —C(═S)—;

L¹ may be —(CH₂)_(m)—, and m may be selected from the group consistingof integers from 0 to 2;

wherein 0 or 1 methylene unit of L¹ may be replaced by —C(O)— or—C(═S)—.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —S— or —(R²)N—;

X may be -L³-L²-;

wherein L² may be —C(O)—;

L³ may be —(CH₂)_(n)—, and n may be 4 or 5;

wherein 1 methylene unit of L³ may be replaced by —NR⁸—, —O—, —S— or—SO—;

R² may be a C₁₋₆ aliphatic group.

For example, the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —S— or —(R²)N—;

X may be -L³-L²-;

wherein L² may be —C(O)—;

L³ may be —(CH₂)_(n)—, and n may be 4 or 5;

wherein 1 methylene unit of L³ may be replaced by —O—;

R² may be a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —O—;

X may be -L³-L²-;

wherein L² may be —C(O)—;

L³ may be —(C(R^(7a))(R^(7b)))_(n)—, and n may be 4 or 5;

wherein 1 methylene unit of L³ may be replaced by —NR⁸— or —O—, and 0 or1 methylene unit of L³ may also be independently replaced by —C(O)— or—C(═S)—;

wherein each R^(7a), each R^(7b) and each R⁸ may each independently behydrogen or a C₁₋₆ aliphatic group.

For example, the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —O—;

X may be -L³-L²-;

wherein L² may be —C(O)—;

L³ may be —(C(R^(7a))(R^(7b)))₄—;

wherein 1 methylene unit of L³ may be replaced by —NR⁸— or —O—;

wherein each R^(7a), each R^(7b) and each R⁸ may each independently behydrogen or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —O—;

X may be -L³-L²-;

wherein L² may be —C(O)—;

L³ may be —(C(R^(7a))(R^(7b)))₄—;

wherein 1 methylene unit of L³ may be replaced by —NR⁸—;

wherein each R^(7a), each R^(7b) and each R⁸ may each independently behydrogen or a C₁₋₆ aliphatic group.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-A):

wherein, R¹ may be —NH—;

X may be -L³-L²-;

wherein L² may be —C(O)—;

L³ may be —(CH₂)_(n)—, and n may be 4 or 5;

wherein 1 methylene unit of L³ may be replaced by —S—.

In one embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise the following group ofstructures:

wherein R² may be a C₁₋₆ aliphatic group which may be optionallyreplaced by R, wherein R may be hydrogen, protium, deuterium, tritium,halogen, —NO₂, —CN, —OH, —SH, —NH₂, —C(O)H, —CO₂H, —C(O)C(O)H,—C(O)CH₂C(O)H, —S(O)H, —S(O)₂H, —C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H,or a C₁₋₆ aliphatic group, or wherein, R² may be halogen, —NO₂, —CN,—OH, —SH, —NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H,—S(O)₂H, —C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphaticgroup.

For example, R² may be methyl which may be optionally substituted withone or more hydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN,—OH, —SH, —NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H,—S(O)₂H, —C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or C₁₋₆ aliphatic groups.For example, R² may be ethyl which may be optionally substituted withhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group. Forexample, R² may be propyl which may be optionally substituted withhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group.

In a second aspect, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(I-B):

wherein, X^(a) may be nitrogen generated by removal of two hydrogenatoms from an amino group of the cytotoxic drug;

L may be -L_(a)-L_(b)-L_(c)-;

-L_(a)- may be selected from the group consisting of:

wherein W may be —(C(R^(wa))(R^(wb)))_(wn)—, Y may be—(OCH₂CH₂)_(yn)—O_(yp)—, and Z may be —(C(R^(za))(R^(zb))_(zn);

wherein wn may be selected from the group consisting of integers ≥0, and

0 or no less than 1 methylene unit of W may be independently replaced by-Cyr-, —N(R^(wx))C(O)—, —C(O)N(R^(wx))—, —C(O)—, —OC(O)—, —C(O)O—,—NR^(wx)—, —O—, —S—, —SO—, —SO₂—, —P(R^(wx))—, —P(═O)(R^(wx))—,—N(R_(wx) ⁾SO₂—, —SO₂N(R^(wx))—, —C(═S)—, —C(═N^(Rwx))—, —N═N—, —C═N—,—N═C— or —C(═N₂)—;

wherein yn may be selected from the group consisting of integers ≥0, andyp may be 0 or 1;

wherein zn may be selected from the group consisting of integers ≥0, and

0 or no less than 1 methylene unit of Z may be independently replaced by-Cyr-, —N(R^(zx))C(O)—, —C(O)N(R^(zx))—, —C(O)—, —OC(O)—, —C(O)O—,—NR^(zx)—, —O—, —S—, —SO—, —SO₂—, —P(R^(zx))—, —P(═O)(R^(zx))—,—N(R^(zx))SO₂—, —SO₂N(R^(zx))—, —C(═S)—, —C(═NR^(zx))—, —N═N—, —C═N—,—N═C— or —C(═N₂)—;

-Cyr- may be selected from the group consisting of: 6-10 memberedarylene, 5-8 membered heteroarylene, 3-10 membered heterocyclylene, and3-10 membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or may be independently substituted with no lessthan 1 substituent R^(cx);

wherein each R^(wa), each R^(wb), each R^(za), each R^(zb), each R^(wx),each R^(zx) and each R^(cx) may each independently be hydrogen, protium,deuterium, tritium, halogen, —NO₂, —CN, —OR^(r), —SR^(r),—N(R^(ra))(R^(rb)), —C(O)R^(r), —CO₂R^(r), —C(O)C(O)R^(r),—C(O)CH₂C(O)R^(r), —S(O)R^(r), —S(O)₂R^(r), —C(O)N(R^(ra))(R^(rb)),—SO₂N(R^(ra))(R^(rb)), —OC(O)R^(r), —N(R)SO₂R^(r) or a C₁₋₆ aliphaticgroup which may be optionally substituted with R^(r);

wherein each R^(r), each R^(ra) and each R^(rb) may each independentlybe hydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

-L_(b)- represents a peptide residue consisting of 2 to 7 amino acids;

-L_(c)- may be selected from the group consisting of:

wherein R^(L1) and R^(L2) may each independently be selected from thegroup consisting of: hydrogen, protium, deuterium, tritium, halogen,—NO₂, —CN, —OH, —SH, —NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H,—S(O)H, —S(O)₂H, —C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H and a C₁₋₆aliphatic group;

R¹, L¹ and L² are defined as in any formula (I-A) in embodiments of thefirst aspect.

In another embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(II-Bx) or formula (II-By):

wherein, X^(a) may be nitrogen generated by removal of two hydrogenatoms from an amino group of the cytotoxic drug;

L may be -L_(a)-L_(b)-L_(c)-;

-L_(a)- may be selected from the group consisting of:

wherein W may be —(C(R^(wa))(R^(wb)))_(wn)—, Y may be—(OCH₂CH₂)_(yn)—O_(yp)—, and Z may be —(C(R^(za))(R^(zb)))_(zn);

wherein wn may be selected from the group consisting of integers ≥0, and

0 or no less than 1 methylene unit of W may be independently replaced by-Cyr-, —N(R^(wx))C(O)—, —C(O)N(R^(wx))—, —C(O)—, —OC(O)—, —C(O)O—,—NR^(wx)—, —O—, —S—, —SO—, —SO₂—, —P(R^(wx))—, —P(═O)(R^(wx))—,—N(R^(wx))SO₂—, —SO₂N(R^(wx))—, —C(═S)—, —C(═N^(Rwx))—, —N═C— or—C(═N₂)—;

wherein yn may be selected from the group consisting of integers ≥0, andyp may be 0 or 1;

wherein zn may be selected from the group consisting of integers ≥0, and

0 or no less than 1 methylene unit of Z may be independently replaced by-Cyr-, —N(R^(zx))C(O)—, —C(O)N(R^(zx))—, —C(O)—, —OC(O)—, —C(O)O—,—NR^(zx)—, —O—, —S—, —SO—, —SO₂—, —P(R^(zx))—, —P(═O)(R^(zx))—,—N(R^(zx))SO₂—, —SO₂N(R^(zx))—, —C(═S)—, —C(═NR^(zx))—, —N═C— or—C(═N₂)—;

-Cyr- may be selected from the group consisting of: 6-10 memberedarylene, 5-8 membered heteroarylene, 3-10 membered heterocyclylene, and3-10 membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or may be independently substituted with no lessthan 1 substituent R^(cx);

wherein each R^(wa), each R^(wb), each R^(za), each R^(zb), each R^(wx),each R^(zx) and each R_(cx) may each independently be hydrogen, protium,deuterium, tritium, halogen, —NO₂, —CN, —OR^(r), —SR^(r), —N(R″—C(O)R^(r), —CO₂R^(r), —C(O)C(O)R^(r), —C(O)CH₂C(O)R^(r), —S(O)R^(r),—S(O)₂R^(r), —C(O)N(R^(ra))(R^(rb)), —SO₂N(R^(ra))(R^(rb)), —OC(O)R^(r),—N(R)SO₂R^(r) or a C₁₋₆ aliphatic group which may be optionallysubstituted with R^(r);

wherein each R^(r), each R^(ra) and each R^(rb) may each independentlybe hydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

-L_(b)- represents a peptide residue consisting of 2 to 7 amino acids;

-L_(c)- may be selected from the group consisting of:

wherein R^(L1) and R^(L2) may each independently be selected from thegroup consisting of: hydrogen, protium, deuterium, tritium, halogen,—NO₂, —CN, —OH, —SH, —NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H,—S(O)H, —S(O)₂H, —C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H and a C₁₋₆aliphatic group;

L², p, ring A, X¹ and L¹ are defined as in any formula (II-Ax) inembodiments of the first aspect;

or X², q, ring A, X¹ and L¹ are defined as in any formula (II-Ay) inembodiments of the first aspect.

In another embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-B):

wherein, X^(a) may be nitrogen generated by removal of two hydrogenatoms from an amino group of the cytotoxic drug;

L may be -L_(a)-L_(b)-L_(c)-;

-L_(a)- may be selected from the group consisting of:

wherein W may be —(C(R^(wa))(R^(wb)))_(wn)-, Y may be—(OCH₂CH₂)_(yn)—O_(yp), and Z may be —(C(R^(za))(R^(zb)))_(zn);

wherein wn may be selected from the group consisting of integers ≥0, and

0 or no less than 1 methylene unit of W may be independently replaced by-Cyr-, —N(R^(wx))C(O)—, —C(O)N(R^(wx))—, —C(O)—, —OC(O)—, —C(O)O—,—NR^(wx)—, —O—, —S—, —SO—, —SO₂—, —P(R^(wx))—, —P(═O)(R^(wx))—,—N(R^(wx))SO₂—, —SO₂N(R^(wx))—, —C(═S)—, —C(═NR^(wx))—, —N═N—, —C═N—,—N═C— or —C(═N₂)—;

wherein yn may be selected from the group consisting of integers ≥0, andyp may be 0 or 1;

wherein zn may be selected from the group consisting of integers ≥0, and

0 or no less than 1 methylene unit of Z may be independently replaced by-Cyr-, —N(R^(zx))C(O)—, —C(O)N(R^(zx))—, —C(O)—, —OC(O)—, —C(O)O—,—NR^(zx)—, —O—, —S—, —SO—, —SO₂—, —P(R^(zx))—, —P(═O)(R^(zx))—,—N(R^(zx))SO₂—, —SO₂N(R^(zx))—, —C(═S)—, —C(═NR^(zx))—, —N═N—, —C═N—,—N═C— or —C(═N₂)—;

-Cyr- may be selected from the group consisting of: 6-10 memberedarylene, 5-8 membered heteroarylene, 3-10 membered heterocyclylene, and3-10 membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or may be independently substituted with no lessthan 1 stituent R^(cx);

wherein each R^(wa), each R^(wb), each R^(za), each R^(zb), each R^(wx),each R^(zx) and each R^(cx) may each independently be hydrogen, protium,deuterium, tritium, halogen, —NO₂, —CN, —OR^(r), —SR^(r),—N(R^(ra))(R^(rb)), —C(O)R^(r), —CO₂R^(r), —C(O)C(O)R^(r),—C(O)CH₂C(O)R^(r), —S(O)R^(r), —S(O)₂R^(r), —C(O)N(R^(ra))(R^(rb)),—SO₂N(R^(ra))(R^(rb)), —OC(O)R^(r), —N(R)SO₂R^(r) or a C₁₋₆ aliphaticgroup which may be optionally substituted with R^(r);

wherein each R^(r), each R^(ra) and each R^(rb) may each independentlybe hydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

-L_(b)- represents a peptide residue consisting of 2 to 7 amino acids;

-L_(c)- may be selected from the group consisting of:

wherein R^(L1) and R^(L2) may each independently be selected from thegroup consisting of: hydrogen, protium, deuterium, tritium, halogen,—NO₂, —CN, —OH, —SH, —NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H,—S(O)H, —S(O)₂H, —C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H and a C₁₋₆aliphatic group;

wherein R¹ and X are defined as in any formula (III-A) in embodiments ofthe first aspect.

In another embodiment, wn may be selected from the group consisting ofintegers from 2 to 6, and 0 or 1 methylene unit of W may beindependently replaced by -Cyr-, —N(R^(wx))C(O)—, —C(O)N(R^(wx))—,—C(O)—, —NR^(wx)— or —O—.

For example, wn may be 1, 2, 3 or 6, and 1 methylene unit of W may beindependently replaced by -Cyr-, —N(R^(wx))C(O)—, —C(O)N(R^(wx))— or—C(O)—.

In another embodiment, yn may be selected from the group consisting ofintegers from 0 to 12, and yp may be 0 or 1.

For example, yn may be 0, 4 or 8, and yp may be 0 or 1.

In another embodiment, zn may be selected from the group consisting ofintegers from 0 to 10, and 0 or 1 methylene unit of Z may beindependently replaced by -Cyr-, —N(R^(zx))C(O)—, —C(O)N(R^(zx))—or—C(O)—.

For example, zn may be 1, 2 or 3, and 1 methylene unit of Z may beindependently replaced by -Cyr-, —N(R^(zx))C(O)—, —C(O)N(R^(zx))— or—C(O)—.

In another embodiment, -Cyr- may be selected from the group consistingof: 6-10 membered arylene and 3-10 membered saturated or partiallyunsaturated carbocyclylene, wherein -Cyr- is unsubstituted or may beindependently substituted with 1 to 3 substituent R^(cx).

For example, -Cyr- may be 3-10 membered saturated carbocyclylene,wherein -Cyr- is unsubstituted or may be independently substituted with1 to 3 substituent R^(cx).

In another embodiment, each R^(wa), each R^(wb), each R^(za), eachR^(zb), each R^(wx), each R^(zx) each R^(cx) may each independently behydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OR^(r),—SR^(r), —N(R^(ra))(R^(rb)), —C(O)R^(r), —CO₂R^(r), —C(O)C(O)R^(r),—C(O)CH₂C(O)R^(r), —S(O)R^(r), —S(O)₂R^(r), —C(O)N(R^(ra))(R^(rb)),—SO₂N(R^(ra))(R^(rb)), —OC(O)R^(r), —N(R)SO₂R^(r), or a C₁₋₆ aliphaticgroup which may be optionally substituted with R^(r); each R^(r), eachR^(ra) and each R^(rb) may each independently be hydrogen, protium,deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH, —NH₂, —C(O)H, —CO₂H,—C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H, —C(O)NH₂, —SO₂NH₂, —OC(O)H,—N(H)SO₂H or a C₁₋₆ aliphatic group.

For example, each R^(wa), each R^(wb), each R^(za), each R^(zb), eachR^(wx), each R^(zx) and each R^(cx) may each independently be hydrogen,halogen, —OR^(r), or a C₁₋₆ aliphatic group which may be optionallysubstituted with R^(r); each R^(r) may independently be hydrogen,halogen or a C₁₋₆ aliphatic group.

In another embodiment, -L_(b)- represents a peptide residue consistingof 2 to 7 amino acids, and the peptide residue of -L_(b)- may be apeptide residue which may be formed of amino acids which may be selectedfrom the group consisting of: phenylalanine, glycine, alanine, valine,citrulline, lysine, serine, glutamic acid and aspartic acid.

For example, -L_(b)- represents a peptide residue consisting of 2 to 4amino acids, and the peptide residue of -L_(b)- may be a peptide residuewhich may be formed of amino acids which may be selected from the groupconsisting of: phenylalanine, glycine, alanine, valine, citrulline andlysine.

For example, -L_(b)- may be selected from the group consisting of:

For example, -L_(b)- may be

In another embodiment, -L_(a)- may be selected from the group consistingof:

For example, -L_(a)- may be selected from the group consisting of:

-L_(c)- may be

In another embodiment, R^(L1) and R^(L2) may each independently beselected from the group consisting of: hydrogen, protium, deuterium,tritium, halogen, —NO₂, —CN, —OH, —SH, —NH₂, —C(O)H, —CO₂H, —C(O)C(O)H,—C(O)CH₂C(O)H, —S(O)H, —S(O)₂H, —C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂Hand a C₁₋₆ aliphatic group.

For example, R^(L1) and R^(L2) may each independently be selected fromthe group consisting of: hydrogen, halogen, —OH and a C₁₋₆ aliphaticgroup.

In another embodiment, -L_(a)- may be

In another embodiment, -L_(b)- may be selected from the group consistingof:

For example, -L_(b)- may be selected from the group consisting of:

In one embodiment, -L_(c)- may be

In one embodiment, -L_(a)-L_(b)-L_(c)- may be selected from the groupconsisting of:

The cytotoxic drug is shown as formula (EXA):

In a third aspect, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(I-C):

wherein, L may be -L_(a)-L_(b)-L_(c)-, and L_(a), L_(b) and L_(c) aredefined as in any formula (I-B) in embodiments of the second aspect;

R¹, L¹ and L² are defined as in any formula (I-A) in embodiments of thefirst aspect.

In another embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(II-Cx) or formula (II-Cy):

wherein, L may be -L_(a)-L_(b)-L_(c)-, and L_(a), L_(b) and L_(c) aredefined as in any formula (II-Bx) in embodiments of the second aspect;

L², p, ring A, X¹ and L¹ are defined as in any formula (II-Ax) inembodiments of the first aspect;

or X², q, ring A, X¹ and L¹ are defined as in any formula (II-Ay) inembodiments of the first aspect.

In another embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-C):

wherein, L may be -L_(a)-L_(b)-L_(c)-, and L_(a), L_(b) and L_(c) aredefined as in any formula (III-B) in embodiments of the second aspect;

wherein R¹ and X are defined as in any formula (III-A) in embodiments ofthe first aspect.

In a fourth aspect, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(I-D):

wherein, Ab may be a ligand, and an average connection number N^(a) maybe an integer or a decimal from 1 to 10;

L may be -L_(a)-L_(b)-L_(c)-, and L_(a), L_(b) and L_(c) are defined asin any formula (I-B) in embodiments of the second aspect;

R¹, L¹ and L² are defined as in any formula (I-A) in embodiments of thefirst aspect.

In another embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(II-Dx) or formula (II-Dy):

wherein, Ab may be a ligand, and an average connection number N^(a) maybe an integer or a decimal from 1 to 10;

L may be -L_(a)-L_(b)-L_(c)-, and L_(a), L_(b) and L_(c) are defined asin any formula (II-Bx) in embodiments of the second aspect;

L², ring A, X¹ and L¹ are defined as in any formula (II-Ax) inembodiments of the first aspect;

or X², ring A, X¹ and L¹ are defined as in any formula (II-Ay) inembodiments of the first aspect.

In another embodiment, the present application provides a compound or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, wherein the compound may comprise a structure shown as formula(III-D):

wherein, Ab may be a ligand, and an average connection number N^(a) maybe an integer or a decimal from 1 to 10;

L may be -L_(a)-L_(b)-L_(c)-, and L_(a), L_(b) and L_(c) are defined asin any formula (III-B) in embodiments of the second aspect;

wherein R¹ and X are defined as in any formula (III-A) in embodiments ofthe first aspect.

In another embodiment, the ligand Ab may be an antibody or anantigen-binding fragment thereof.

For example, the ligand Ab may be selected from the group consisting of:a chimeric antibody, a humanized antibody and a fully humanizedantibody.

For example, the ligand Ab targets the following: HER2, HER3, B7H3,TROP2, Claudin 18.2, CD30, CD33, CD70 and EGFR.

For example, the ligand Ab targets the following; for example, theantibody may be an antibody that targets the following target points:5T4, AGS-16, ANGPTL4, ApoE, CD19, CTGF, CXCR5, FGF2, MCPT8, MFI2, MS4A7,NCA, Sema5b, SLITRK6, STC2, TGF, 0772P, 5T4, ACTA2, ADGRE1, AG-7, AIF1,AKR1C1, AKR1C2, ASLG659, Axl, B7H3, BAFF-R, BCMA, BMPR1B, BNIP3, C1QA,C1QB, CA6, CADM1, CCD79b, CCL5, CCR5, CCR7, CD11c, CD123, CD138, CD142,CD147, CD166, CD19, CD19, CD22, CD21, CD20, CD205, CD22, CD223, CD228,CD25, CD30, CD33, CD37, CD38, CD40, CD45, CD45 (PTPRC), CD46, CD47,CD49D (ITGA4), CD56, CD66e, CD70, CD71, CD72, CD74, CD79a, CD79b, CD80,CDCP1, CDH11, CD11b, CEA, CEACAM5, c-Met, COL6A3, COL7A1, CRIPTO, CSF1R,CTSD, CTSS, CXCL11, CXCL10, DDIT4, DLL3, DLL4, DR5, E16, EFNA4, EGFR,EGFRvIII, EGLN, EGLN3, EMR2, ENPP3, EpCAM, EphA2, EphB2R, ETBR, FcRH2,FcRH1, FGFR2, FGFR3, FLT3, FOLR-α, GD2, GEDA, GPC-1, GPNMB, GPR20, GZMB,HER2, HER3, HLA-DOB, HMOX1, IFI6, IFNG, IGF-1R, IGFBP3, IL10RA1, IL-13R,IL-2, IL20Ra, IL-3, IL-4, IRTA2, KISS1R, KRT33A, LIV-1, LOX, LRP-1,LRRC15, LUM, LY64, LY6E, Ly86, LYPD3, MDP, MMP10, MMP14, MMP16, MPF,MSG783, MSLN, MUC-1, NaPi2b, Napi3b, Nectin-4, Nectin-4, NOG, P2X5,pCAD, P-Cadherin, PDGFRA, PDK1, PD-L¹, PFKFB3, PGF, PGK1, PIK3AP1,PIK3CD, PLOD2, PSCA, PSCAh1g, PSMA, PSMA, PTK7, P-Cadherin, RNF43,NaPi2b, ROR1, ROR2, SERPINE1, SLC39A6, SLTRK6, STAT1, STEAP1, STEAP2,TCF4, TENB2, TGFB1, TGFB2, TGFBR1, TNFRSF21, TNFSF9, Trop-2, TrpM4,Tyrol, UPK1B, VEGFA, WNT5A, epidermal growth factors, brevican,mesothelin, sodium phosphate cotransporter 2B, Claudin 18.2, endothelinreceptors, mucins (such as mucin 1 and mucin 16), guanylate cyclase C,integrin a4p7, integrin a5p6, trophoblast glycoprotein, and tissuefactors.

In another embodiment, the average connection number N^(a) may be aninteger or a decimal from 2 to 8. For example, the average connectionnumber N^(a) may be an integer or a decimal from 3 to 8. For example,the average connection number N^(a) may be an integer or a decimal from1 to 2, 2 to 3, 3 to 4, 4 to 5, 5 to 6, 6 to 7, 7 to 8, 8 to 9, or 9 to10.

In a fifth aspect, the present application provides a compound ofgeneral formula (I-E) or a tautomer, a mesomer, a racemate, anenantiomer or a diastereoisomer thereof, or a mixture thereof, or apharmaceutically acceptable salt thereof,

wherein R¹, L¹ and L² are defined as in any formula (I-A) in embodimentsof the first aspect.

In another embodiment, the present application provides a compound ofgeneral formula (II-E_(x)) or (II-E_(y)), or a tautomer, a mesomer, aracemate, an enantiomer or a diastereoisomer thereof, or a mixturethereof, or a pharmaceutically acceptable salt thereof,

wherein L², p, ring A, X¹ and L¹ are defined as in any formula (II-Ax)in embodiments of the first aspect;

or X², q, ring A, X¹ and L¹ are defined as in any formula (II-Ay) inembodiments of the first aspect.

In another embodiment, the present application provides a compound ofgeneral formula (M-E) or a tautomer, a mesomer, a racemate, anenantiomer or a diastereoisomer thereof, or a mixture thereof, or apharmaceutically acceptable salt thereof,

wherein R¹ and X are defined as in any formula (III-A) in embodiments ofthe first aspect.

In a sixth aspect, the present application provides a compound ofgeneral formula (I-F) or a tautomer, a mesomer, a racemate, anenantiomer or a diastereoisomer thereof, or a mixture thereof, or apharmaceutically acceptable salt thereof,

wherein, L^(x) may be -L_(ax)-L_(b)-L_(c)-;

-L_(ax)- may be selected from the group consisting of:

wherein R^(hal) may be iodine or bromine;

wherein W may be —(C(R^(wa))(R^(wb)))_(wn)—, Y may be—(OCH₂CH₂)_(yn)—O_(yp)—, and Z may be —(C(R^(za))(R^(zb)))_(zn);

wherein wn may be selected from the group consisting of integers ≥0, and

0 or no less than 1 methylene unit of W may be independently replaced by-Cyr-, —N(R^(wx))C(O)—, —C(O)N(R^(wx))—, —C(O)—, —OC(O)—, —C(O)O—,—NR^(wx)—, —O—, —S—, —SO—, —SO₂—, —P(R^(wx))—, —P(═O)(R^(wx))—,—N(R^(wx))SO₂—, —SO₂N(R^(wx))—, —C(═S)—, —C(═N^(Rwx))—, —N═C— or—C(═N₂)—;

wherein yn may be selected from the group consisting of integers ≥0, andyp may be 0 or 1;

wherein zn may be selected from the group consisting of integers ≥0, and

0 or no less than 1 methylene unit of Z may be independently replaced by-Cyr-, —N(R^(zx))C(O)—, —C(O)N(R^(zx))—, —C(O)—, —OC(O)—, —C(O)O—,—NR^(zx)—, —O—, —S—, —SO—, —SO₂—, —P(R^(zx))—, —P(═O)(R^(zx))—,—N(R^(zx))SO₂—, —SO₂N(R^(zx))—, —C(═S)—, —C(═NR^(zx))—, —N═C— or—C(═N₂)—;

-Cyr- may be selected from the group consisting of: 6-10 memberedarylene, 5-8 membered heteroarylene, 3-10 membered heterocyclylene, and3-10 membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or may be independently substituted with no lessthan 1 substituent R^(cx);

wherein each R^(wa), each R^(wb), each R^(za), each R^(zb), each R^(wx),each R^(zx) and each R^(cx) may each independently be hydrogen, protium,deuterium, tritium, halogen, —NO₂, —CN, —OR^(r), —SR^(r),—N(R^(ra))(R^(rb)), —C(O)R^(r), —CO₂R^(r), —C(O)C(O)R^(r),—C(O)CH₂C(O)R^(r), —S(O)R^(r), —S(O)₂R^(r), —C(O)N(R^(ra))(R^(rb)),—SO₂N(R^(ra))(R^(rb)), —OC(O)R^(r), —N(R)SO₂R^(r) or a C₁₋₆ aliphaticgroup which may be optionally substituted with R^(r);

wherein each R^(r), each R^(ra) and each R^(rb) may each independentlybe hydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

L^(b) and L^(c) are defined as in any formula (I-B) in embodiments ofthe second aspect;

R¹, L¹ and L² are defined as in any formula (I-A) in embodiments of thefirst aspect.

In another embodiment, the present application provides a compound ofgeneral formula (II-F_(x)) or (II-F_(y)), or a tautomer, a mesomer, aracemate, an enantiomer or a diastereoisomer thereof, or a mixturethereof, or a pharmaceutically acceptable salt thereof,

wherein, L^(x) may be -L_(ax)-L_(b)-L_(c)-;

-L_(ax)- may be selected from the group consisting of:

wherein R^(hal) may be iodine or bromine;

wherein W may be —(C(R^(wa))(R^(wb)))_(wn)—, Y may be—(OCH₂CH₂)_(yn)—O_(yp)—, and Z may be —(C(R^(za))(R^(zb)))_(zn);

wherein wn may be selected from the group consisting of integers ≥0, and

0 or no less than 1 methylene unit of W may be independently replaced by-Cyr-, —N(R^(wx))C(O)—, —C(O)N(R^(wx))—, —C(O)—, —OC(O)—, —C(O)O—,—NR^(wx)—, —O—, —S—, —SO—, —SO₂—, —P(R^(wx))—, —P(═O)(R^(wx))—,—N(R^(wx))SO₂—, —SO₂N(R^(wx))—, —C(═S)—, —C(═NR^(wx))—, —N═C— or—C(═N₂)—;

wherein yn may be selected from the group consisting of integers ≥0, andyp may be 0 or 1;

wherein zn may be selected from the group consisting of integers ≥0, and

0 or no less than 1 methylene unit of Z may be independently replaced by-Cyr-, —N(R^(zx))C(O)—, —C(O)N(R^(zx))—, —C(O)—, —OC(O)—, —C(O)O—,—NR^(zx)—, —O—, —S—, —SO—, —SO₂—, —P(R^(zx))—, —P(═O)(R^(zx))—,—N(R^(zx))SO₂—, —SO₂N(R^(zx))—, —C(═S)—, —C(═NR^(zx))—, —N═C— or—C(═N₂)—;

-Cyr- may be selected from the group consisting of: 6-10 memberedarylene, 5-8 membered heteroarylene, 3-10 membered heterocyclylene, and3-10 membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or may be independently substituted with no lessthan 1 substituent R^(cx);

wherein each R^(wa), each R^(wb), each R^(za), each R^(zb), each R^(wx),each R^(zx) and each R^(cx) may each independently be hydrogen, protium,deuterium, tritium, halogen, —NO₂, —CN, —OR^(r), —SR^(r),—N(R^(ra))(R^(rb)), —C(O)R^(r), —CO₂R^(r), —C(O)C(O)R^(r),—C(O)CH₂C(O)R^(r), —S(O)R^(r), —S(O)₂R^(r), —C(O)N(R^(ra))(R^(rb)),—SO₂N(R^(ra))(R^(rb)), —OC(O)R^(r), —N(R)SO₂R^(r) or a C₁₋₆ aliphaticgroup which may be optionally substituted with R^(r);

wherein each R^(r), each R^(ra) and each R^(rb) may each independentlybe hydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

L^(b) and L^(c) are defined as in any formula (II-Bx) in embodiments ofthe second aspect;

wherein L², p, ring A, X¹ and L¹ are defined as in any formula (II-Ax)in embodiments of the first aspect;

or X², q, ring A, X¹ and L¹ are defined as in any formula (II-Ay) inembodiments of the first aspect.

In another embodiment, the present application provides a compound ofgeneral formula (III-F) or a tautomer, a mesomer, a racemate, anenantiomer or a diastereoisomer thereof, or a mixture thereof, or apharmaceutically acceptable salt thereof,

wherein, L^(x) may be -L_(ax)-L_(b)-L_(c)-;

-L_(ax)- may be selected from the group consisting of:

wherein R^(hal) may be iodine or bromine;

wherein W may be —(C(R^(wa))(R^(wb)))_(wn)—, Y may be—(OCH₂CH₂)_(yn)—O_(yp)—, and Z may be —(C(R^(za))(R^(zb)))_(zn);

wherein wn may be selected from the group consisting of integers ≥0, and

0 or no less than 1 methylene unit of W may be independently replaced by-Cyr-, —N(R^(wx))C(O)—, —C(O)N(R^(wx))—, —C(O)—, —OC(O)—, —C(O)O—,—NR^(wx)—, —O—, —S—, —SO—, —SO₂—, —P(R^(wx))—, —P(═O)(R^(wx))—,—N(R^(wx))SO₂—, —SO₂N(R^(wx))—, —C(═S)—, —C(═NR^(wx))—, —N═C— or—C(═N₂)—;

wherein yn may be selected from the group consisting of integers ≥0, andyp may be 0 or 1;

wherein zn may be selected from the group consisting of integers ≥0, and

0 or no less than 1 methylene unit of Z may be independently replaced by-Cyr-, —N(R^(zx))C(O)—, —C(O)N(R^(zx))—, —C(O)—, —OC(O)—, —C(O)O—,—NR^(zx)—, —O—, —S—, —SO—, —SO₂—, —P(R^(zx))—, —P(═O)(R^(zx))—,—N(R^(zx))SO₂—, —SO₂N(R^(zx))—, —C(═S)—, —C(═NR^(zx))—, —N═C— or—C(═N₂)—;

-Cyr- may be selected from the group consisting of: 6-10 memberedarylene, 5-8 membered heteroarylene, 3-10 membered heterocyclylene, and3-10 membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or may be independently substituted with no lessthan 1 substituent R^(cx);

wherein each R^(wa), each R^(wb), each R^(za), each R^(zb), each R^(wx),each R^(zx) and each R^(cx) may each independently be hydrogen, protium,deuterium, tritium, halogen, —NO₂, —CN, —OR^(r), —SR^(r),—N(R^(ra))(R^(rb)), —C(O)R^(r), —CO₂R^(r), —C(O)C(O)R^(r),—C(O)CH₂C(O)R^(r), —S(O)R^(r), —S(O)₂R^(r), —C(O)N(R^(ra))(R^(rb)),—SO₂N(R^(ra))(R^(rb)), —OC(O)R^(r), —N(R)SO₂R^(r) or a C₁₋₆ aliphaticgroup which may be optionally substituted with R^(r);

wherein each R^(r), each R^(ra) and each R^(rb) may each independentlybe hydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

L^(b) and L^(c) are defined as in any formula (III-B) in embodiments ofthe second aspect;

wherein R¹ and X are defined as in any formula (III-A) in embodiments ofthe first aspect.

In another embodiment, L_(ax)- may be selected from the group consistingof:

wherein R^(hal) may be iodine or bromine;

wherein W may be —(C(R^(wa))(R^(wb)))_(wn)—, Y may be—(OCH₂CH₂)_(yn)—O_(yp)—, and Z may be —(C(R^(za))(R^(zb)))_(zn).

In another embodiment, wn may be selected from the group consisting ofintegers from 2 to 6, and 0 or 1 methylene unit of W may beindependently replaced by -Cyr-, —N(R^(wx))C(O)—, —C(O)N(R^(wx))—,—C(O)—, —NR^(wx)— or —O—.

For example, wn may be 1, 2, 3 or 6, and 1 methylene unit of W may beindependently replaced by -Cyr-, —N(R^(wx))C(O)—, —C(O)N(R^(wx))— or—C(O)—.

In another embodiment, yn may be selected from the group consisting ofintegers from 0 to 12, and yp may be 0 or 1.

For example, yn may be 0, 4 or 8, and yp may be 0 or 1.

In another embodiment, zn may be selected from the group consisting ofintegers from 0 to 10, and 0 or 1 methylene unit of Z may beindependently replaced by -Cyr-, —N(R^(zx))C(O)—, —C(O)N(R^(zx))—or—C(O)—.

For example, zn may be 1, 2 or 3, and 1 methylene unit of Z may beindependently replaced by -Cyr-, —N(R^(zx))C(O)—, —C(O)N(R^(zx))—or—C(O)—.

In another embodiment, -Cyr- may be selected from the group consistingof: 6-10 membered arylene and 3-10 membered saturated or partiallyunsaturated carbocyclylene, wherein -Cyr- is unsubstituted or may beindependently substituted with 1 to 3 substituent R^(cx).

For example, -Cyr- may be 3-10 membered saturated carbocyclylene,wherein -Cyr- is unsubstituted or may be independently substituted with1 to 3 substituent R^(cx).

In another embodiment, each R^(wa), each R^(wb), each R^(za), eachR^(zb), each R^(wx), each R^(zx) and each R^(cx) may each independentlybe hydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OR^(r),—SR^(r), —N(R^(ra))(R^(rb)), —C(O)R^(r), —CO₂R^(r), —C(O)C(O)R^(r),—C(O)CH₂C(O)R^(r), —S(O)R^(r), —S(O)₂R^(r), —C(O)N(R^(ra))(R^(rb)),—SO₂N(R^(ra))(R^(rb)), —OC(O)R^(r), —N(R)SO₂R^(r), or a C₁₋₆ aliphaticgroup which may be optionally substituted with R^(r); each R^(r), eachR^(ra) and each R^(rb) may each independently be hydrogen, protium,deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH, —NH₂, —C(O)H, —CO₂H,—C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H, —C(O)NH₂, —SO₂NH₂, —OC(O)H,—N(H)SO₂H or a C₁₋₆ aliphatic group.

For example, each R^(wa), each R^(wb), each R^(za), each R^(zb), eachR^(wx), each R^(zx) and each R^(cx) may each independently be hydrogen,halogen, —OR^(r), or a C₁₋₆ aliphatic group which may be optionallysubstituted with R^(r); each R^(r) may independently be hydrogen,halogen or a C₁₋₆ aliphatic group.

In another embodiment, L_(ax)- may be

In another embodiment, L_(ax)-L_(b)-L_(c)- may be selected from thegroup consisting of:

Compounds Disclosed Herein

In one embodiment, the compounds disclosed herein include, but are notlimited to:

No. Structure P-I-1

P-I-2

P-I-3

P-I-4

P-I-5

P-I-6

P-I-7

P-I-8

P-I-9

P-I-10

P-I-11

P-I-12

P-I-13

P-I-14

P-I-15

P-I-16

P-I-17

P-I-18

P-I-19

P-I-20

P-I-21

P-I-22

P-I-23

P-I-24

P-I-25

P-I-26

P-I-27

P-I-28

P-I-29

P-I-30

P-I-31

P-I-32

P-I-33

P-I-34

P-I-35

P-I-36

P-I-37

P-I-38

P-I-39

P-I-40

P-I-41

P-I-42

P-I-43

P-I-44

P-I-45

P-I-46

P-I-47

L-I-1

L-I-2

L-I-3

L-I-4

L-I-5

L-I-6

L-I-7

L-I-8

L-I-9

L-I-10

L-I-11

L-I-12

L-I-13

L-I-14

L-I-15

L-I-16

L-I-17

L-I-18

L-I-19

L-I-20

L-I-21

L-I-22

L-I-23

L-I-24

L-I-25

L-I-26

L-I-27

L-I-28

L-I-29

L-I-30

L-I-31

L-I-32

L-I-33

L-I-34

L-I-35

L-I-36

L-I-37

L-I-38

L-I-39

L-I-40

L-I-41

L-I-42

L-I-43

L-I-44

L-I-45

L-I-46

L-I-47

L-I-48

L-I-49

L-I-50

ADC-I-1

ADC-I-2

ADC-I-3

ADC-I-4

ADC-I-5

ADC-I-6

ADC-I-7

ADC-I-8

ADC-I-9

ADC-I-10

ADC-I-11

ADC-I-12

ADC-I-13

ADC-I-14

ADC-I-15

ADC-I-16

ADC-I-17

ADC-I-18

ADC-I-19

ADC-I-20

ADC-I-21

ADC-I-22

ADC-I-23

ADC-I-24

ADC-I-25

ADC-I-26

ADC-I-27

ADC-I-28

ADC-I-29

ADC-I-30

The average connection number n in the above list may be an integer or adecimal from 1 to 10. The average connection number n in the above listmay be an integer or a decimal from 2 to 8. For example, the averageconnection number n may be an integer or a decimal from 3 to 8. Forexample, the average connection number n may be an integer or a decimalfrom 1 to 2, 2 to 3, 3 to 4, 4 to 5, 5 to 6, 6 to 7, 7 to 8, 8 to 9, or9 to 10.

No. Structure P-II- 1

P-II- 2

P-II- 3

P-II- 4

P-II- 5

P-II- 6

P-II- 7

P-II- 8

P-II- 9

P-II- 10

P-II- 11

P-II- 12

P-II- 13

P-II- 14

P-II- 15

P-II- 16

P-II- 17

P-II- 18

P-II- 19

P-II- 20

P-II- 21

P-II- 22

P-II- 23

P-II- 24

P-II- 25

L-II- 1

L-II- 2

L-II- 3

L-II- 4

L-II- 5

L-II- 6

L-II- 7

L-II- 8

L-II- 9

L-II- 10

L-II- 11

L-II- 12

L-II- 13

L-II- 14

L-II- 15

L-II- 16

L-II- 17

L-II- 18

L-II- 19

L-II- 20

L-II- 21

L-II- 22

L-II- 23

L-II- 24

L-II- 25

L-II- 26

L-II- 27

L-II- 28

ADC- II-1

ADC- II-2

ADC- II-3

ADC- II-4

ADC- II-5

ADC- II-6

ADC- II-7

ADC- II-8

ADC- II-9

ADC- II-10

ADC- II-11

ADC- II-12

ADC- II-13

ADC- II-14

ADC- II-15

ADC- II-16

ADC- II-17

ADC- II-18

ADC- II-19

ADC- II-20

ADC- II-21

ADC- II-22

ADC- II-23

ADC- II-24

ADC- II-25

ADC- II-26

ADC- II-27

ADC- II-28

ADC- II-29

ADC- II-30

ADC- II-31

ADC- II-32

ADC- II-33

ADC- II-34

ADC- II-35

ADC- II-36

ADC- II-37

ADC- II-38

ADC- II-39

ADC- II-40

ADC- II-41

ADC- II-42

ADC- II-43

ADC- II-44

ADC- II-45

ADC- II-46

ADC- II-47

ADC- II-48

ADC- II-49

ADC- II-50

ADC- II-51

ADC- II-52

ADC- II-53

ADC- II-54

ADC- II-55

ADC- II-56

ADC- II-57

ADC- II-58

The average connection number n in the above list may be an integer or adecimal from 1 to 10. The average connection number n in the above listmay be an integer or a decimal from 2 to 8. For example, the averageconnection number n may be an integer or a decimal from 3 to 8. Forexample, the average connection number n may be an integer or a decimalfrom 1 to 2, 2 to 3, 3 to 4, 4 to 5, 5 to 6, 6 to 7, 7 to 8, 8 to 9, or9 to 10.

In one embodiment, the compounds disclosed herein include, but are notlimited to:

No. Structure P-III-1

P-III-2

P-III-3

P-III-4

P-III-5

P-III-6

P-III-7

P-III-8

P-III-9

P-III-10

P-III-11

P-III-12

P-III-13

P-III-14

P-III-15

P-III-16

P-III-17

P-III-18

P-III-19

P-III-20

P-III-21

P-III-22

P-III-23

P-III-24

P-III-25

P-III-26

P-III-27

P-III-28

P-III-29

P-III-30

P-III-31

L-III-1

L-III-2

L-III-3

L-III-4

L-III-5

L-III-6

L-III-7

L-III-8

L-III-9

L-III-10

L-III-11

L-III-12

L-III-13

L-III-14

L-III-15

L-III-16

L-III-17

L-III-18

L-III-19

L-III-20

L-III-21

L-III-22

L-III-23

L-III-24

L-III-25

L-III-26

L-III-27

L-III-28

L-III-29

L-III-30

L-III-31

ADC-III-1

ADC-III-2

ADC-III-3

ADC-III-4

ADC-III-5

ADC-III-6

ADC-III-7

ADC-III-8

ADC-III-9

ADC-III-10

ADC-III-11

ADC-III-12

ADC-III-13

ADC-III-14

ADC-III-15

ADC-III-16

ADC-III-17

ADC-III-18

ADC-III-19

ADC-III-20

ADC-III-21

ADC-III-22

ADC-III-23

ADC-III-24

ADC-III-25

ADC-III-26

ADC-III-27

ADC-III-28

ADC-III-29

ADC-III-30

ADC-III-31

ADC-III-32

ADC-III-33

ADC-III-34

ADC-III-35

The average connection number n in the above list may be an integer or adecimal from 1 to 10. The average connection number n in the above listmay be an integer or a decimal from 2 to 8. For example, the averageconnection number n may be an integer or a decimal from 3 to 8. Forexample, the average connection number n may be an integer or a decimalfrom 1 to 2, 2 to 3, 3 to 4, 4 to 5, 5 to 6, 6 to 7, 7 to 8, 8 to 9, or9 to 10.

Ligands

The ligands described herein may be protein hormones, lectin, growthfactors, antibodies, or other molecules capable of binding to a cell, areceptor and/or an antigen. For example, the ligand disclosed herein maybe an antibody or an antigen-binding fragment thereof.

In the present application, the ligand may comprise at least one CDR inthe light chain variable region VL of an antibody. The CDRs may bedefined according to Kabat.

In the present application, the antigen-binding protein may comprise anLCDR1, and the LCDR1 may comprise an amino acid sequence set forth inany one of SEQ ID NOs: 1-4. The CDRs may be defined according to Kabat.

In the present application, the antigen-binding protein may comprise anLCDR2, and the LCDR2 may comprise an amino acid sequence set forth inany one of SEQ ID NOs: 5-8. The CDRs may be defined according to Kabat.

In the present application, the antigen-binding protein may comprise anLCDR3, and the LCDR3 may comprise an amino acid sequence set forth inany one of SEQ ID NOs: 9-12. The CDRs may be defined according to Kabat.

In the present application, the isolated antigen-binding protein maycomprise LCDRs 1-3, wherein the LCDR1 may comprise an amino acidsequence set forth in any one of SEQ ID NOs: 1-4, the LCDR2 may comprisean amino acid sequence set forth in any one of SEQ ID NOs: 5-8, and theLCDR3 may comprise an amino acid sequence set forth in any one of SEQ IDNOs: 9-12. The CDRs may be defined according to Kabat.

For example, the antigen-binding protein described herein may comprisethe same LCDRs 1-3 as trastuzumab, wherein the LCDR1 may comprise anamino acid sequence set forth in SEQ ID NO: 1, the LCDR2 may comprise anamino acid sequence set forth in SEQ ID NO: 5, and the LCDR3 maycomprise an amino acid sequence set forth in SEQ ID NO: 9. For example,the antigen-binding protein described herein may comprise the same LCDRs1-3 as pertuzumab, wherein the LCDR1 may comprise an amino acid sequenceset forth in SEQ ID NO: 2, the LCDR2 may comprise an amino acid sequenceset forth in SEQ ID NO: 6, and the LCDR3 may comprise an amino acidsequence set forth in SEQ ID NO: 10. The CDRs may be defined accordingto Kabat.

For example, the antigen-binding protein described herein may comprisethe same LCDRs 1-3 as sacituzumab, wherein the LCDR1 may comprise anamino acid sequence set forth in SEQ ID NO: 3, the LCDR2 may comprise anamino acid sequence set forth in SEQ ID NO: 7, and the LCDR3 maycomprise an amino acid sequence set forth in SEQ ID NO: 11. The CDRs maybe defined according to Kabat.

For example, the antigen-binding protein described herein may comprisethe same LCDRs 1-3 as zolbetuximab, wherein the LCDR1 may comprise anamino acid sequence set forth in SEQ ID NO: 4, the LCDR2 may comprise anamino acid sequence set forth in SEQ ID NO: 8, and the LCDR3 maycomprise an amino acid sequence set forth in SEQ ID NO: 12. The CDRs maybe defined according to Kabat.

The antigen-binding protein described herein may comprise at least oneCDR in the heavy chain variable region VH of an antibody. The CDRs maybe defined according to Kabat.

In the present application, the antigen-binding protein may comprise anHCDR1, and the HCDR1 may comprise an amino acid sequence set forth inany one of SEQ ID NOs: 13-16. The CDRs may be defined according toKabat.

In the present application, the antigen-binding protein may comprise anHCDR2, and the HCDR2 may comprise an amino acid sequence set forth inany one of SEQ ID NOs: 17-20. The CDRs may be defined according toKabat.

In the present application, the antigen-binding protein may comprise anHCDR3, and the HCDR3 may comprise an amino acid sequence set forth inany one of SEQ ID NOs: 21-24. The CDRs may be defined according toKabat.

In the present application, the isolated antigen-binding protein maycomprise HCDRs 1-3, wherein the HCDR1 may comprise an amino acidsequence set forth in any one of SEQ ID NOs: 13-16, the HCDR2 maycomprise an amino acid sequence set forth in any one of SEQ ID NOs:17-20, and the HCDR3 may comprise an amino acid sequence set forth inany one of SEQ ID NOs: 21-24. The CDRs may be defined according toKabat.

For example, the antigen-binding protein described herein may comprisethe same HCDRs 1-3 as trastuzumab, wherein the HCDR1 may comprise anamino acid sequence set forth in SEQ ID NO: 13, the HCDR2 may comprisean amino acid sequence set forth in SEQ ID NO: 17, and the HCDR3 maycomprise an amino acid sequence set forth in SEQ ID NO: 21. The CDRs maybe defined according to Kabat.

For example, the antigen-binding protein described herein may comprisethe same HCDRs 1-3 as pertuzumab, wherein the HCDR1 may comprise anamino acid sequence set forth in SEQ ID NO: 14, the HCDR2 may comprisean amino acid sequence set forth in SEQ ID NO: 18, and the HCDR3 maycomprise an amino acid sequence set forth in SEQ ID NO: 22. The CDRs maybe defined according to Kabat.

For example, the antigen-binding protein described herein may comprisethe same HCDRs 1-3 as sacituzumab, wherein the HCDR1 may comprise anamino acid sequence set forth in SEQ ID NO: 15, the HCDR2 may comprisean amino acid sequence set forth in SEQ ID NO: 19, and the HCDR3 maycomprise an amino acid sequence set forth in SEQ ID NO: 23. The CDRs maybe defined according to Kabat.

For example, the antigen-binding protein described herein may comprisethe same HCDRs 1-3 as zolbetuximab, wherein the HCDR1 may comprise anamino acid sequence set forth in SEQ ID NO: 16, the HCDR2 may comprisean amino acid sequence set forth in SEQ ID NO: 20, and the HCDR3 maycomprise an amino acid sequence set forth in SEQ ID NO: 24. The CDRs maybe defined according to Kabat.

In the present application, the isolated antigen-binding protein maycomprise LCDRs 1-3 and HCDRs 1-3, wherein The LCDR1 may comprise anamino acid sequence set forth in any one of SEQ ID NOs: 1-4, the LCDR2may comprise an amino acid sequence set forth in any one of SEQ ID NOs:5-8, the LCDR3 may comprise an amino acid sequence set forth in any oneof SEQ ID NOs: 9-12, the HCDR1 may comprise an amino acid sequence setforth in any one of SEQ ID NOs: 13-16, the HCDR2 may comprise an aminoacid sequence set forth in any one of SEQ ID NOs: 17-20, and the HCDR3may comprise an amino acid sequence set forth in any one of SEQ ID NOs:21-24. The CDRs may be defined according to Kabat.

For example, the antigen-binding protein described herein may comprisethe same LCDRs 1-3 and HCDRs 1-3 as trastuzumab, wherein the LCDR1 maycomprise an amino acid sequence set forth in SEQ ID NO: 1, the LCDR2 maycomprise an amino acid sequence set forth in SEQ ID NO: 5, the LCDR3 maycomprise an amino acid sequence set forth in SEQ ID NO: 9, the HCDR1 maycomprise an amino acid sequence set forth in SEQ ID NO: 13, the HCDR2may comprise an amino acid sequence set forth in SEQ ID NO: 17, and theHCDR3 may comprise an amino acid sequence set forth in SEQ ID NO: 21.The CDRs may be defined according to Kabat.

For example, the antigen-binding protein described herein may comprisethe same LCDRs 1-3 and HCDRs 1-3 as pertuzumab, wherein the LCDR1 maycomprise an amino acid sequence set forth in SEQ ID NO: 2, the LCDR2 maycomprise an amino acid sequence set forth in SEQ ID NO: 6, the LCDR3 maycomprise an amino acid sequence set forth in SEQ ID NO: 10, the HCDR1may comprise an amino acid sequence set forth in SEQ ID NO: 14, theHCDR2 may comprise an amino acid sequence set forth in SEQ ID NO: 18,and the HCDR3 may comprise an amino acid sequence set forth in SEQ IDNO: 22. The CDRs may be defined according to Kabat.

For example, the antigen-binding protein described herein may comprisethe same LCDRs 1-3 and HCDRs 1-3 as sacituzumab, wherein the LCDR1 maycomprise an amino acid sequence set forth in SEQ ID NO: 3, the LCDR2 maycomprise an amino acid sequence set forth in SEQ ID NO: 7, the LCDR3 maycomprise an amino acid sequence set forth in SEQ ID NO: 11, the HCDR1may comprise an amino acid sequence set forth in SEQ ID NO: 15, theHCDR2 may comprise an amino acid sequence set forth in SEQ ID NO: 19,and the HCDR3 may comprise an amino acid sequence set forth in SEQ IDNO: 23. The CDRs may be defined according to Kabat.

For example, the antigen-binding protein described herein may comprisethe same LCDRs 1-3 and HCDRs 1-3 as zolbetuximab, wherein the LCDR1 maycomprise an amino acid sequence set forth in SEQ ID NO: 4, the LCDR2 maycomprise an amino acid sequence set forth in SEQ ID NO: 8, the LCDR3 maycomprise an amino acid sequence set forth in SEQ ID NO: 12, the HCDR1may comprise an amino acid sequence set forth in SEQ ID NO: 16, theHCDR2 may comprise an amino acid sequence set forth in SEQ ID NO: 20,and the HCDR3 may comprise an amino acid sequence set forth in SEQ IDNO: 24. The CDRs may be defined according to Kabat.

In the present application, the antigen-binding protein may comprise alight chain variable region VL, and the VL may comprise an amino acidsequence set forth in any one of SEQ ID NOs: 25-28.

In the present application, the antigen-binding protein may comprise aheavy chain variable region VH, and the VH may comprise an amino acidsequence set forth in any one of SEQ ID NOs: 29-32.

In the present application, the antigen-binding protein may comprise alight chain variable region VL and a heavy chain variable region VH,wherein the VL may comprise an amino acid sequence set forth in any oneof SEQ ID NOs: 25-28, and the VH may comprise an amino acid sequence setforth in any one of SEQ ID NOs: 29-32.

For example, the antigen-binding protein described herein may comprisethe same light chain variable region VL and heavy chain variable regionVH as trastuzumab, wherein the VL may comprise an amino acid sequenceset forth in SEQ ID NO: 25, and the VH may comprise an amino acidsequence set forth in SEQ ID NO: 29.

For example, the antigen-binding protein described herein may comprisethe same light chain variable region VL and heavy chain variable regionVH as pertuzumab, wherein the VL may comprise an amino acid sequence setforth in SEQ ID NO: 26, and the VH may comprise an amino acid sequenceset forth in SEQ ID NO: 30.

For example, the antigen-binding protein described herein may comprisethe same light chain variable region VL and heavy chain variable regionVH as sacituzumab, wherein the VL may comprise an amino acid sequenceset forth in SEQ ID NO: 27, and the VH may comprise an amino acidsequence set forth in SEQ ID NO: 31.

For example, the antigen-binding protein described herein may comprisethe same light chain variable region VL and heavy chain variable regionVH as zolbetuximab, wherein the VL may comprise an amino acid sequenceset forth in SEQ ID NO: 28, and the VH may comprise an amino acidsequence set forth in SEQ ID NO: 32.

In the present application, the antigen-binding protein may comprise alight chain, and the light chain may comprise an amino acid sequence setforth in any one of SEQ ID NOs: 33-36.

In the present application, the antigen-binding protein may comprise aheavy chain, and the heavy chain may comprise an amino acid sequence setforth in any one of SEQ ID NOs: 37-40.

In the present application, the antigen-binding protein may comprise anantibody light chain and an antibody heavy chain, wherein the lightchain may comprise an amino acid sequence set forth in any one of SEQ IDNOs: 33-36, and the heavy chain may comprise an amino acid sequence setforth in any one of SEQ ID NOs: 37-40.

For example, the antigen-binding protein described herein may comprisethe same antibody light chain and antibody heavy chain as trastuzumab,wherein the light chain may comprise an amino acid sequence set forth inSEQ ID NO: 33, and the heavy chain may comprise an amino acid sequenceset forth in SEQ ID NO: 37.

For example, the antigen-binding protein described herein may comprisethe same antibody light chain and antibody heavy chain as pertuzumab,wherein the light chain may comprise an amino acid sequence set forth inSEQ ID NO: 34, and the heavy chain may comprise an amino acid sequenceset forth in SEQ ID NO: 38.

For example, the antigen-binding protein described herein may comprisethe same antibody light chain and antibody heavy chain as sacituzumab,wherein the light chain may comprise an amino acid sequence set forth inSEQ ID NO: 35, and the heavy chain may comprise an amino acid sequenceset forth in SEQ ID NO: 39.

For example, the antigen-binding protein described herein may comprisethe same antibody light chain and antibody heavy chain as zolbetuximab,wherein the light chain may comprise an amino acid sequence set forth inSEQ ID NO: 36, and the heavy chain may comprise an amino acid sequenceset forth in SEQ ID NO: 40.

Prevention and/or Prevention of Tumors

In another aspect, the present application provides use of the compoundor the tautomer, the mesomer, the racemate, the enantiomer or thediastereoisomer thereof, or the mixture thereof, or the pharmaceuticallyacceptable salt thereof, or the pharmaceutical composition thereofdescribed herein, in preparing a medicament for treating and/orpreventing a tumor. The tumor may be selected from the group consistingof tumors associated with expression of the following: HER2, HER3, B7H3,TROP2, Claudin 18.2, CD30, CD33, CD70 and EGFR. The tumor may beselected from the group consisting of: lung cancer, kidney cancer,urinary tract carcinoma, colorectal cancer, prostatic cancer,glioblastoma multiforme, ovarian cancer, pancreatic cancer, breastcancer, melanoma, liver cancer, bladder cancer, stomach cancer andesophageal cancer.

In another aspect, the present application provides a method fortreating and/or preventing a tumor, which comprises administering to asubject in need the compound or the tautomer, the mesomer, the racemate,the enantiomer or the diastereoisomer thereof, or the mixture thereof,or the pharmaceutically acceptable salt thereof described herein, and/orthe pharmaceutical composition that may comprise the same. The tumor maybe selected from the group consisting of tumors associated withexpression of the following: HER2, HER3, B7H3, TROP2, Claudin 18.2,CD30, CD33, CD70 and EGFR. The tumor may be selected from the groupconsisting of: lung cancer, kidney cancer, urinary tract carcinoma,colorectal cancer, prostatic cancer, glioblastoma multiforme, ovariancancer, pancreatic cancer, breast cancer, melanoma, liver cancer,bladder cancer, stomach cancer and esophageal cancer.

In another aspect, the present application provides the compound or thetautomer, the mesomer, the racemate, the enantiomer or thediastereoisomer thereof, or the mixture thereof, or the pharmaceuticallyacceptable salt thereof described herein, and/or the pharmaceuticalcomposition that may comprise the same, for use in treating and/orpreventing a tumor. The tumor may be selected from the group consistingof tumors associated with expression of the following: HER2, HER3, B7H3,TROP2, Claudin 18.2, CD30, CD33, CD70 and EGFR. The tumor may beselected from the group consisting of: lung cancer, kidney cancer,urinary tract carcinoma, colorectal cancer, prostatic cancer,glioblastoma multiforme, ovarian cancer, pancreatic cancer, breastcancer, melanoma, liver cancer, bladder cancer, stomach cancer andesophageal cancer.

For example, the tumor may be selected from the group consisting oftumors associated with expression of the following: 5T4, AGS-16,ANGPTL4, ApoE, CD19, CTGF, CXCR5, FGF2, MCPT8, MFI2, MS4A7, NCA, Sema5b,SLITRK6, STC2, TGF, 0772P, 5T4, ACTA2, ADGRE1, AG-7, AWL AKR1C1, AKR1C2,ASLG659, Axl, B7H3, BAFF-R, BCMA, BMPR1B, BNIP3, C1QA, C1QB, CA6, CADM1,CCD79b, CCL5, CCR5, CCR7, CD11c, CD123, CD138, CD142, CD147, CD166,CD19, CD19, CD22, CD21, CD20, CD205, CD22, CD223, CD228, CD25, CD30,CD33, CD37, CD38, CD40, CD45, CD45 (PTPRC), CD46, CD47, CD49D (ITGA4),CD56, CD66e, CD70, CD71, CD72, CD74, CD79a, CD79b, CD80, CDCP1, CDH11,CD1 1b, CEA, CEACAM5, c-Met, COL6A3, COL7A1, CRIPTO, CSF1R, CTSD, CTSS,CXCL11, CXCL10, DDIT4, DLL3, DLL4, DR5, E16, EFNA4, EGFR, EGFRvIII,EGLN, EGLN3, EMR2, ENPP3, EpCAM, EphA2, EphB2R, ETBR, FcRH2, FcRH1,FGFR2, FGFR3, FLT3, FOLR-a, GD2, GEDA, GPC-1, GPNMB, GPR20, GZMB, HER2,HER3, HLA-DOB, HMOX1, IFI6, IFNG, IGF-1R, IGFBP3, IL10RA1, IL-13R, IL-2,IL20Ra, IL-3, IL-4, IL-6, IRTA2, KISS1R, KRT33A, LIV-1, LOX, LRP-1,LRRC15, LUM, LY64, LY6E, Ly86, LYPD3, MDP, MMP10, MMP14, MMP16, MPF,MSG783, MSLN, MUC-1, NaPi2b, Napi3b, Nectin-4, Nectin-4, NOG, P2X5,pCAD, P-Cadherin, PDGFRA, PDK1, PD-L¹, PFKFB3, PGF, PGK1, PIK3AP1,PIK3CD, PLOD2, PSCA, PSCAhlg, PSMA, PSMA, PTK7, P-Cadherin, RNF43,NaPi2b, ROR1, ROR2, SERPINE1, SLC39A6, SLTRK6, STAT1, STEAP1, STEAP2,TCF4, TENB2, TGFB1, TGFB2, TGFBR1, TNFRSF21, TNFSF9, Trop-2, TrpM4,Tyrol, UPK1B, VEGFA, WNT5A, epidermal growth factors, brevican,mesothelin, sodium phosphate cotransporter 2B, Claudin 18.2, endothelinreceptors, mucins (such as mucin 1 and mucin 16), guanylate cyclase C,integrin a4p7, integrin a5p6, trophoblast glycoprotein, and tissuefactors.

The compound described herein may have inhibitory activity against invitro proliferation of tumor cells. The inhibitory activity may be that:compared with in a culture medium of tumor cells to which a negativecontrol or a control drug is added, the proliferation capacity of thetumor cells is reduced by no less than 1%, no less than 2%, no less than4%, no less than 5%, no less than 8%, no less than 10%, no less than15%, no less than 18%, no less than 20%, no less than 25%, no less than40%, no less than 50%, no less than 60%, no less than 70%, no less than80%, no less than 90% or no less than 95% in a culture medium to whichthe compound disclosed herein is added. For example, the inhibitoryactivity may be an IC₅₀ value (nM) for tumor cells of no more than10000, no more than 5000, no more than 4000, no more than 3000, no morethan 2000, no more than 1000, no more than 500, no more than 400, nomore than 300, no more than 200, no more than 150, no more than 120, nomore than 110, no more than 100, no more than 99, no more than 98, nomore than 97, no more than 95, no more than 90, no more than 80, no morethan 75, no more than 70, no more than 65, no more than 62, no more than60, no more than 50, no more than 40, no more than 30, no more than 25,no more than 23, no more than 22, no more than 20, no more than 19, nomore than 18, no more than 18.5, no more than 17, no more than 15, nomore than 12, no more than 10, no more than 9, no more than 8.5, no morethan 7, no more than 6.7, no more than 6, no more than 5.9, no more than5.5, no more than 5.0, no more than 4.8, no more than 4.5, no more than4.4, no more than 4, no more than 3.5, no more than 3, no more than 2.5,no more than 2, no more than 1.5, no more than 1.0, no more than 0.5, nomore than 0.3, no more than 0.29, no more than 0.25, no more than 0.21,no more than 0.20, no more than 0.18, no more than 0.17, no more than0.15, no more than 0.12, no more than 0.10, no more than 0.09, no morethan 0.08, no more than 0.07, no more than 0.06, no more than 0.05, nomore than 0.04, no more than 0.03, no more than 0.02 or no more than0.01. For example, the tumor cells may include, but are not limited to,solid tumor cells; for example, the tumor cells include, but are notlimited to, gastric cancer cells, or breast cancer cells; for example,the tumor cells may include, but are not limited to, NCI-N87 cells,JIMT-1 cells or MBA-MB-231 cells.

The compound described herein may have targeting inhibition. Thetargeting inhibition may be that: compared with in a culture medium oftumor cells with high expression of a specific target point to which anegative control or a control drug is added, the proliferation capacityof the tumor cells with high expression of a specific target point isreduced by no less than 1%, no less than 2%, no less than 4%, no lessthan 5%, no less than 8%, no less than 10%, no less than 15%, no lessthan 18%, no less than 20%, no less than 25%, no less than 40%, no lessthan 50%, no less than 60%, no less than 70%, no less than 80%, no lessthan 90% or no less than 95% in a culture medium to which the compounddisclosed herein is added. For example, the targeting inhibition may bean IC₅₀ value (nM), for tumor cells with high expression of a specifictarget point, of no more than 10000, no more than 5000, no more than4000, no more than 3000, no more than 2000, no more than 1000, no morethan 500, no more than 400, no more than 300, no more than 200, no morethan 185, no more than 150, no more than 120, no more than 110, no morethan 100, no more than 99, no more than 98, no more than 97, no morethan 95, no more than 91, no more than 80, no more than 74, no more than70, no more than 65, no more than 62, no more than 60, no more than 50,no more than 40, no more than 30, no more than 25, no more than 23, nomore than 22, no more than 20, no more than 19, no more than 18, no morethan 18.5, no more than 17, no more than 15, no more than 12, no morethan 10, no more than 9, no more than 8.5, no more than 7, no more than6.7, no more than 6, no more than 5.9, no more than 5.5, no more than5.0, no more than 4.8, no more than 4.5, no more than 4.4, no more than4, no more than 3.5, no more than 3, no more than 2.5, no more than 2,no more than 1.5, no more than 1.0, no more than 0.5, no more than 0.3,no more than 0.29, no more than 0.25, no more than 0.21, no more than0.20, no more than 0.18, no more than 0.17, no more than 0.15, no morethan 0.12, no more than 0.10, no more than 0.09, no more than 0.08, nomore than 0.07, no more than 0.06, no more than 0.05, no more than 0.04,no more than 0.03, no more than 0.02 or no more than 0.01. For example,the tumor cells with high expression of a specific target point mayinclude, but are not limited to, solid tumor cells; for example, thetumor cells with high expression of a specific target point include, butare not limited to, gastric cancer cells, or breast cancer cells; forexample, the tumor cells with high expression of a specific target pointmay include, but are not limited to, NCI-N87 cells or JIMT-1 cells. Thespecific target point may include, but is not limited to, HER2 or TROP2.

The compound described herein may have plasma stability. The plasmastability may be that: the compound disclosed herein releases no morethan 50%, no more than 40%, no more than 30%, no more than 20%, no morethan 10%, no more than 7%, no more than 5%, no more than 4%, no morethan 3%, no more than 2%, no more than 1.9%, no more than 1.8%, no morethan 1.7%, no more than 1.6%, no more than 1.5%, no more than 1.4%, nomore than 1.3%, no more than 1.2%, no more than 1.1%, no more than 1.0%,no more than 0.9%, no more than 0.8%, no more than 0.7%, no more than0.6%, no more than 0.5%, no more than 0.4%, no more than 0.3%, no morethan 0.2% or no more than 0.1% of the cytotoxic drug 1 day, 3 days, 5days, 7 days, 14 days, 20 days or 30 days after the compound is added toplasma.

The compound described herein may have in vivo tumor-inhibiting effect.The tumor-inhibiting effect may be that: compared with the case where anegative control or a control drug is administered to an animal, thetumor of the animal is reduced in volume by no less than 1%, no lessthan 2%, no less than 4%, no less than 5%, no less than 8%, no less than10%, no less than 15%, no less than 18%, no less than 20%, no less than25%, no less than 40%, no less than 50%, no less than 55%, no less than60%, no less than 70%, no less than 73%, no less than 75%, no less than80%, no less than 90% or no less than 95% 1 day, 3 days, 5 days, 7 days,14 days, 20 days, 21 days or 30 days after the compound disclosed hereinis administered, or the tumor of the animal is reduced in volume by noless than 1.1 fold, no less than 1.3 fold, no less than 1.5 fold, noless than 2 fold, no less than 3 fold, no less than 5 fold, no less than10 fold, no less than 20 fold, no less than 22 fold, no less than 30fold, no less than 50 fold, no less than 100 fold, no less than 500fold, no less than 1000 fold or no less than 1500 fold 1 day, 3 days, 5days, 7 days, 14 days, 20 days, 21 days or 30 days after the compounddisclosed herein is administered. The animal may include, but is notlimited to, a mammal. For example, the animal may include, but is notlimited to, a cat, a dog, a horse, a pig, a cow, a sheep, a rabbit, amouse, a rat, a monkey or a human. The administration may include, butis not limited to, oral administration, intravenous injection,intravenous drip, intraperitoneal injection or topical administration.

The compound described herein may have a bystander effect. The bystandereffect may be that: the compound disclosed herein has no obviousinhibiting effect against cell proliferation of the tumor cells with lowexpression of a specific target point, but in the co-culturing of thetumor cells with low expression of the specific target point and thetumor cells with high expression of the specific target point, thecompound disclosed herein can simultaneously inhibit the cellproliferation of the tumor cells with low expression of the specifictarget point and the tumor cells with high expression of the specifictarget point. For example, in the co-culturing of the tumor cells withlow expression of the specific target point and the tumor cells withhigh expression of the specific target point, the inhibiting activitymay be an IC₅₀ value (nM), for the tumor cells with low expression ofthe specific target point, of no more than 10000, no more than 5000, nomore than 4000, no more than 3000, no more than 2000, no more than 1000,no more than 500, no more than 400, no more than 300, no more than 200,no more than 185, no more than 150, no more than 120, no more than 110,no more than 100, no more than 99, no more than 98, no more than 97, nomore than 95, no more than 91, no more than 80, no more than 74, no morethan 70, no more than 65, no more than 62, no more than 60, no more than50, no more than 40, no more than 30, no more than 25, no more than 23,no more than 22, no more than 20, no more than 19, no more than 18, nomore than 18.5, no more than 17, no more than 15, no more than 12, nomore than 10, no more than 9, no more than 8.5, no more than 7, no morethan 6.7, no more than 6, no more than 5.9, no more than 5.5, no morethan 5.0, no more than 4.8, no more than 4.5, no more than 4.4, no morethan 4, no more than 3.5, no more than 3, no more than 2.5, no more than2, no more than 1.5, no more than 1.0, no more than 0.5, no more than0.3, no more than 0.29, no more than 0.25, no more than 0.21, no morethan 0.20, no more than 0.18, no more than 0.17, no more than 0.15, nomore than 0.12, no more than 0.10, no more than 0.09, no more than 0.08,no more than 0.07, no more than 0.06, no more than 0.05, no more than0.04, no more than 0.03, no more than 0.02 or no more than 0.01.Compared with in tumor cells with high expression of the specific targetpoint, the expression of the specific target point in tumor cells withlow expression of the specific target point may be reduced by no lessthan 1%, no less than 2%, no less than 4%, no less than 5%, no less than8%, no less than 10%, no less than 15%, no less than 18%, no less than20%, no less than 25%, no less than 40%, no less than 50%, no less than60%, no less than 70%, no less than 80%, no less than 90% or no lessthan 95%. For example, the tumor cells with high expression of aspecific target point may include, but are not limited to, solid tumorcells; for example, the tumor cells with high expression of a specifictarget point include, but are not limited to, gastric cancer cells, orbreast cancer cells; for example, the tumor cells with high expressionof a specific target point may include, but are not limited to, NCI-N87cells or JIMT-1 cells. For example, the tumor cells with low expressionof a specific target point may include, but are not limited to, solidtumor cells; for example, the tumor cells with low expression of aspecific target point include, but are not limited to, breast cancercells; for example, the tumor cells with low expression of a specifictarget point may include, but are not limited to, HCC1187 cells.

The compound described herein may have capacity in inhibiting transportvia a transporter. The capacity in inhibiting transport may be areduction in the efflux ratio of the compound described herein by noless than 1%, no less than 2%, no less than 4%, no less than 5%, no lessthan 8%, no less than 10%, no less than 15%, no less than 18%, no lessthan 20%, no less than 25%, no less than 40%, no less than 50%, no lessthan 60%, no less than 70%, no less than 80%, no less than 90% or noless than 95% compared with a standard of a transport substrate. Forexample, the testing of the efflux ratio may be a method commonly usedby those skilled in the art, or may be described in the examples of thepresent application.

The compound described herein may have in vivo tumor targetingcapability. The in vivo targeting ability may be that: when the compoundlabeled with a signal substance is administered to an animal, comparedwith in other tissues and organs of the animal, the distribution of thelabeled compound in a tumor tissue may be increased by no less than 1%,no less than 2%, no less than 4%, no less than 5%, no less than 8%, noless than 10%, no less than 15%, no less than 18%, no less than 20%, noless than 25%, no less than 40%, no less than 50%, no less than 60%, noless than 70%, no less than 80%, no less than 90% or no less than 95%,or may be increased by no less than 1.1 fold, no less than 1.3 fold, noless than 1.5 fold, no less than 2 fold, no less than 3 fold, no lessthan 5 fold, no less than 10 fold, no less than 20 fold, no less than 22fold, no less than 30 fold, no less than 50 fold, no less than 100 fold,no less than 500 fold, no less than 1000 fold or no less than 1500 fold.The signal substance may be a radioactive material; for example, thesignal substance includes, but is not limited to, ¹²⁵I. The animal mayinclude, but is not limited to, a mammal. For example, the animal mayinclude, but is not limited to, a cat, a dog, a horse, a pig, a cow, asheep, a rabbit, a mouse, a rat, a monkey or a human. The administrationmay include, but is not limited to, oral administration, intravenousinjection, intravenous drip, intraperitoneal injection or topicaladministration. The tissues or organs may include, but are not limitedto, heart, liver, spleen, lung, kidney, brain or bone marrow.

The compound described herein may have good in vivo safety. The in vivosafety may be that: after the compound disclosed herein is administeredto an animal, the release rate of in vivo free toxin in the animal is nomore than 50%, no more than 40%, no more than 30%, no more than 20%, nomore than 10%, no more than 7%, no more than 5%, no more than 4%, nomore than 3%, no more than 2%, no more than 1.9%, no more than 1.8%, nomore than 1.7%, no more than 1.6%, no more than 1.5%, no more than 1.4%,no more than 1.3%, no more than 1.2%, no more than 1.1%, no more than1.0%, no more than 0.9%, no more than 0.8%, no more than 0.7%, no morethan 0.6%, no more than 0.5%, no more than 0.4%, no more than 0.3%, nomore than 0.2% or no more than 0.1%. For example, the in vivo safety maybe that: the compound described herein may be administered at aconcentration of no less than 0.5 mg/kg, no less than 1 mg/kg, no lessthan 2 mg/kg, no less than 3 mg/kg, no less than 4 mg/kg, no less than 5mg/kg, no less than 10 mg/kg, no less than 20 mg/kg, no less than 30mg/kg, no less than 50 mg/kg, no less than 70 mg/kg, no less than 100mg/kg, no less than 200 mg/kg, no less than 500 mg/kg or no less than1000 mg/kg without causing toxic manifestation in the animal. Forexample, the animal may include, but is not limited to, a cat, a dog, ahorse, a pig, a cow, a sheep, a rabbit, a mouse, a rat, a monkey or ahuman. The administration may include, but is not limited to, oraladministration, intravenous injection, intravenous drip, intraperitonealinjection or topical administration.

Pharmaceutical Composition

The pharmaceutical composition described herein may contain, in additionto the active compound, one or more adjuvants, which may be selectedfrom the group consisting of the following ingredients: fillers(diluents), binders, wetting agents, disintegrants, excipients, and thelike. Depending on the method of administration, the composition maycontain 0.1 wt. % to 99% wt. % of the active compound.

The pharmaceutical composition containing the active ingredient may bein a form suitable for oral administration, such as tablet, troche,lozenge, aqueous or oil suspension, dispersible powder or granule,emulsion, hard or soft capsule, or syrup. Oral compositions may beprepared according to any method for preparing pharmaceuticalcompositions known in the art, and the compositions may contain binders,fillers, lubricants, disintegrants, pharmaceutically acceptable wettingagents, and the like, and may also contain one or more ingredients thatmay be selected from the group consisting of: sweetening agents,flavouring agents, coloring agents and preservatives.

Aqueous suspensions may contain the active substance in admixture withexcipients suitable for the formulation of aqueous suspensions. Aqueoussuspensions may also contain one or more preservatives, for example, oneor more coloring agents, one or more flavoring agents, and one or moresweetening agents. Oily suspensions may be formulated by suspending theactive ingredient in a vegetable oil. These oil suspensions may containthickening agents. The sweetening agents and the flavoring agentsdescribed above may also be added.

The pharmaceutical compositions may also be prepared as follows:dispersible powders or granules for preparing aqueous suspensionsprovide the active ingredient, and water is added to mix the activeingredient with one or more of dispersing agents, wetting agents,suspending agents or preservatives. Other excipients, such as sweeteningagents, flavouring agents and coloring agents, may also be added. Thesecompositions are well preserved by the addition of antioxidants such asascorbic acid. The pharmaceutical composition disclosed herein may alsobe in the form of an oil-in-water emulsion.

The pharmaceutical composition may be in the form of a sterileinjectable aqueous solution. Available and acceptable vehicles orsolvents include water, Ringer's solution and isotonic sodium chloridesolution. The sterile injectable formulation may be a sterile injectableoil-in-water microemulsion in which the active ingredient is dissolvedin the oil phase. For example, the active ingredient is dissolved in amixture of soybean oil and lecithin. The oil solution may then be addedto a mixture of water and glycerol and treated to form a microemulsion.The injection or microemulsion can be locally injected into thebloodstream of a patient in large quantities. Alternatively, it may bedesirable to administer solutions and microemulsions in such a way as tomaintain a constant circulating concentration of the compound disclosedherein. To maintain such a constant concentration, a device forcontinuous intravenous drug delivery may be used. For example, thedevice may be a Deltec CADD-PLUS.™. 5400 intravenous injection pump.

The pharmaceutical composition may be in the form of a sterileinjectable aqueous or oily suspension for intramuscular and subcutaneousadministration. The suspension may be prepared according to the knownart using the suitable dispersing agents or wetting agents andsuspending agents described above. The sterile injectable formulationmay also be a sterile injection or suspension prepared in a parenterallyacceptable non-toxic diluent or solvent. Alternatively, a sterile fixedoil may be conveniently used as a solvent or a suspending medium.

The compound disclosed herein may be administered in the form of asuppository for rectal administration. These pharmaceutical compositionsmay be prepared by mixing a drug with a suitable non-irritatingexcipient which may be solid at ordinary temperatures but liquid in therectum and will therefore melt in the rectum to release the drug. Suchmaterials include cocoa butter, glycerinated gelatin, hydrogenatedvegetable oils, and mixtures of polyethylene glycols of variousmolecular weights and fatty acid esters of polyethylene glycol.

As is well known to those skilled in the art, the dosage of the drugadministered depends on a variety of factors, including but not limitedto, the activity of the particular compound employed, the age of thepatient, the weight of the patient, the health condition of the patient,the behavior of the patient, the diet of the patient, the time ofadministration, the mode of administration, the rate of excretion, thecombination of drugs, and the like. In addition, the optimal treatmentregimen, such as the mode of treatment, a compound described herein or atautomer, a mesomer, a racemate, an enantiomer or a diastereoisomerthereof, or a mixture thereof, or a pharmaceutically acceptable saltthereof, and/or the daily amount of the compound or the tautomer, themesomer, the racemate, the enantiomer or the diastereoisomer thereof, orthe mixture thereof, or the pharmaceutically acceptable salt thereof, orthe type of the pharmaceutically acceptable salt thereof, can beverified according to conventional treatment schemes.

Technical Schemes for Synthesis

For the synthesis purpose of the present disclosure, the followingtechnical schemes for synthesis are adopted in the present application:

Step 1: reacting a compound of general formula (Y1) with a compound ofgeneral formula (KI3) in the presence of a condensing agent, optionallyunder a basic condition, to obtain a compound of general formula (I-E-M)

Step 2: removing a protecting group of the compound of general formula(I-E-M) to obtain the compound of general formula (I-E)

wherein,

Rp is a hydroxy protecting group;

R¹, L¹ and L² are defined as in any formula (I-A) in embodiments of thefirst aspect.

Reagents that provide basic conditions include organic bases andinorganic bases, wherein the organic bases include, but are not limitedto, triethylamine, diethylamine, N-methylmorpholine, pyridine,piperidine, NA-diisopropylethylamine, n-butyllithium, lithiumdiisopropylamide, potassium acetate, sodium tert-butoxide, potassiumtert-butoxide, and the like, and the inorganic bases include, but arenot limited to, sodium hydride, potassium carbonate, sodium carbonate,cesium carbonate, sodium hydroxide, lithium hydroxide, and the like.

The condensing agent may be selected from the group consisting of4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride,1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride, N,N′-dicyclohexylcarbodiimide,N,N′-diisopropylcarbodiimide,O-benzotriazol-N,N,N,N-tetramethyluroniumtetrafluoroborate,1-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazol,O-benzotriazol-N,N,N,N-tetramethyluronium hexafluorophosphate,2-(7-azobenzotriazol)-N,N,N,N-tetramethyluronium hexafluorophosphate,benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate,and benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate,preferably 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholiniumchloride, or 1-hydroxybenzotriazole and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.

wherein R¹ is selected from the group consisting of: —O—, —(R²)N—,—P(═O)(R²)— and —S—;

L² is —(C(R^(3a))(R^(3b)))_(m)—R,

wherein 0 or no less than 1 methylene unit of L² is independentlyreplaced by -Cy-, —N(R⁴)C(O)—, —C(O)N(R⁴)—, —C(O)—, —OC(O)—, —C(O)O—,—NR⁴—, —O—, —S—, —SO—, —SO₂—, —P(R⁴)—, —P(═O)(R⁴)—, —N(R⁴)SO₂—,—SO₂N(R⁴)—, —C(═S)—, —C(═NR⁴)—, —N═C— or —C(═N₂)—;

L¹ is —(C(R^(5a))(R^(5b)))_(n)—,

wherein 0 or no less than 1 methylene unit of L¹ is independentlyreplaced by -Cy-, —N(R⁶)C(O)—, —C(O)N(R⁶)—, —C(O)—, —OC(O)—, —C(O)O—,—NR⁶—, —O—, —S—, —SO—, —SO₂—, —P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—,—SO₂N(R⁶)—, —C(═S)—, —C(═NR⁶)—, —N═C— or —C(═N₂)—;

-Cy- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein -Cy-is unsubstituted or independently substituted with no less than 1substituent R⁷;

for example, wherein, R^(3a) and R^(5a) form a ring B together with anatom therebetween, wherein the ring B may be selected from the groupconsisting of: 5-8 membered heteroarylene and 3-10 membered saturated orpartially unsaturated heterocyclylene, and the ring B is unsubstitutedor substituted with no less than 1 substituent R⁸; each R^(3b), each R⁴,each R^(5b) and each R⁶ are each independently hydrogen, protium,deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR, —N(R^(a))(R^(b)),—C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R, —S(O)₂R,—C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆aliphatic group optionally substituted with R; or R^(3a) and R^(5a), R⁴and R^(5a), R^(3a) and R⁶ or R⁴ and R⁶ each independently optionallyform a ring B together with an atom therebetween, wherein the ring B isselected from the group consisting of: 5-8 membered heteroarylene and3-10 membered saturated or partially unsaturated heterocyclylene, andthe ring B is unsubstituted or substituted with no less than 1substituent R⁸;

for example, wherein, R⁴ and R^(5a) form a ring B together with an atomtherebetween, wherein the ring B is selected from the group consistingof: 5-8 membered heteroarylene and 3-10 membered saturated or partiallyunsaturated heterocyclylene, and the ring B is unsubstituted orsubstituted with no less than 1 substituent R⁸; each R^(3a), eachR^(3b), each R^(5b) and each R⁶ are each independently hydrogen,protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R,or a C₁₋₆ aliphatic group optionally substituted with R; or R^(3a) andR^(5a), R⁴ and R^(5a), R^(3a) and R⁶ or R⁴ and R⁶ each independentlyoptionally form a ring B together with an atom therebetween, wherein thering B is selected from the group consisting of: 5-8 memberedheteroarylene and 3-10 membered saturated or partially unsaturatedheterocyclylene, and the ring B is unsubstituted or substituted with noless than 1 substituent R⁸;

for example, wherein, R^(3a) and R⁶ form a ring B together with an atomtherebetween, wherein the ring B is selected from the group consistingof: 5-8 membered heteroarylene and 3-10 membered saturated or partiallyunsaturated heterocyclylene, and the ring B is unsubstituted orsubstituted with no less than 1 substituent R⁸; each R^(3b), each R⁴,each R^(5a) and each R^(5b) are each independently hydrogen, protium,deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR, —N(R^(a))(R^(b)),—C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R, —S(O)₂R,—C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆aliphatic group optionally substituted with R; or R^(3a) and R^(5a), R⁴and R^(5a), R^(3a) and R⁶ or R⁴ and R⁶ each independently optionallyform a ring B together with an atom therebetween, wherein the ring B isselected from the group consisting of: 5-8 membered heteroarylene and3-10 membered saturated or partially unsaturated heterocyclylene, andthe ring B is unsubstituted or substituted with no less than 1substituent R⁸;

for example, wherein, R⁴ and R⁶ independently optionally form a ring Btogether with an atom therebetween, wherein the ring B is selected fromthe group consisting of: 5-8 membered heteroarylene and 3-10 memberedsaturated or partially unsaturated heterocyclylene, and the ring B isunsubstituted or substituted with no less than 1 substituent R⁸; eachR^(3a), each R^(3b), each R^(5a) and each R^(5b) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R,or a C₁₋₆ aliphatic group optionally substituted with R; or R^(3a) andR^(5a), R⁴ and R^(5a), R^(3a) and R⁶ or R⁴ and R⁶ each independentlyoptionally form a ring B together with an atom therebetween, wherein thering B is selected from the group consisting of: 5-8 memberedheteroarylene and 3-10 membered saturated or partially unsaturatedheterocyclylene, and the ring B is unsubstituted or substituted with noless than 1 substituent R⁸;

wherein each R², each R⁷ and each R⁸ are each independently hydrogen,protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R,or a C₁₋₆ aliphatic group optionally substituted with R;

wherein each R, each R^(a) and each R^(b) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

m and n are each independently selected from the group consisting ofintegers ≥1.

Step 1: reacting a compound of general formula (Y1-2) with a compound ofgeneral formula (KI4) in the presence of or in the absence of a reducingagent under an acidic or basic condition to obtain a compound of generalformula (I-E-M)

Step 2: removing a protecting group of the compound of general formula(I-E-M) to obtain the compound of general formula (I-E)

wherein,

Rp is a hydroxy protecting group;

R¹, L¹ and L² are defined as in any formula (I-A) in embodiments of thefirst aspect.

Reducing agents include, but are not limited to, sodium hydride, calciumhydride, lithium hydride, lithium aluminum hydride, sodium borohydride,lithium borohydride, sodium triethylborohydride, sodiumtriacetoxyborohydride and sodium cyanoborohydride.

Reagents that provide acidic conditions include a protic acid and aLewis acid, wherein the protic acid includes, but is not limited to,hydrochloric acid, sulfuric acid, nitric acid, nitrous acid, sulfurousacid, phosphoric acid, phosphorous acid, formic acid, acetic acid,propionic acid, butyric acid, citric acid, benzoic acid,p-toluenesulfonic acid, p-nitrobenzoic acid, methanesulfonic acid,trifluoromethanesulfonic acid and trifluoroacetic acid, and the Lewisacid includes, but is not limited to, boron trifluoride, zinc chloride,magnesium chloride, aluminum chloride, stannic chloride and ferricchloride.

Reagents that provide basic conditions include organic bases andinorganic bases, wherein the organic bases include, but are not limitedto, triethylamine, diethylamine, N-methylmorpholine, pyridine,piperidine, NA-diisopropylethylamine, n-butyllithium, lithiumdiisopropylamide, potassium acetate, sodium tert-butoxide, potassiumtert-butoxide, and the like, and the inorganic bases include, but arenot limited to, sodium hydride, potassium carbonate, sodium carbonate,cesium carbonate, sodium hydroxide, lithium hydroxide, and the like.

The condensing agent may be selected from the group consisting of4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride,1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride, N,N-dicyclohexylcarbodiimide,N,N′-diisopropylcarbodiimide, O-benzotriazol-N,N,N,N-tetramethyluroniumtetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazol,O-benzotriazol-N,N,N,N-tetramethyluronium hexafluorophosphate,2-(7-azobenzotriazol)-N,N,N,N-tetramethyluronium hexafluorophosphate,benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate,and benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate,preferably 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholiniumchloride, or 1-hydroxybenzotriazole and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.

Step 1: reacting a compound of general formula (Y2x) or a compound ofgeneral formula (Y2y) with a compound of formula (KI4) in the presenceof a condensing agent, optionally under a basic condition, to obtain acompound of general formula (II-Ex-M) or general formula (II-Ey-M)

Step 2: removing a protecting group of the compound of general formula(II-Ex-M) or general formula (II-Ey-M) to obtain the compound of generalformula (II-Ex) or general formula (II-Ey)

wherein,

Rp is a hydroxy protecting group;

wherein L², p, ring A, X¹ and L¹ are defined as in any (II-Ax) inembodiments of the first aspect;

or X², q, ring A, X¹ and L¹ are defined as in any (II-Ay) in embodimentsof the first aspect.

Reagents that provide basic conditions include organic bases andinorganic bases, wherein the organic bases include, but are not limitedto, triethylamine, diethylamine, N-methylmorpholine, pyridine,piperidine, NA-diisopropylethylamine, n-butyllithium, lithiumdiisopropylamide, potassium acetate, sodium tert-butoxide, potassiumtert-butoxide, and the like, and the inorganic bases include, but arenot limited to, sodium hydride, potassium carbonate, sodium carbonate,cesium carbonate, sodium hydroxide, lithium hydroxide, and the like.

The condensing agent may be selected from the group consisting of4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride,1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride, N,N-dicyclohexylcarbodiimide,N,N′-diisopropylcarbodiimide, O-benzotriazol-N,N,N,N-tetramethyluroniumtetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazol,O-benzotriazol-N,N,N,N-tetramethyluronium hexafluorophosphate,2-(7-azobenzotriazol)-N,N,N,N-tetramethyluronium hexafluorophosphate,benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate,and benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate,preferably 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholiniumchloride, or 1-hydroxybenzotriazole and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.

reacting a compound of general formula (Y3) with a compound of generalformula (KI4) in the presence of a condensing agent, optionally under abasic condition, to obtain the compound of general formula (III-E-M).

wherein,

R¹ and X are defined as in any (III-A) in embodiments of the firstaspect.

Reagents that provide basic conditions include organic bases andinorganic bases, wherein the organic bases include, but are not limitedto, triethylamine, diethylamine, N-methylmorpholine, pyridine,piperidine, N,N-diisopropylethylamine, n-butyllithium, lithiumdiisopropylamide, potassium acetate, sodium tert-butoxide, potassiumtert-butoxide, and the like, and the inorganic bases include, but arenot limited to, sodium hydride, potassium carbonate, sodium carbonate,cesium carbonate, sodium hydroxide, lithium hydroxide, and the like.

The condensing agent may be selected from the group consisting of4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride,1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride, N,N-dicyclohexylcarbodiimide,N,N′-diisopropylcarbodiimide, O-benzotriazol-N,N,N,N-tetramethyluroniumtetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazol,O-benzotriazol-N,N,N,N-tetramethyluronium hexafluorophosphate,2-(7-azobenzotriazol)-N,N,N,N-tetramethyluronium hexafluorophosphate,benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate,and benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate,preferably 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholiniumchloride, or 1-hydroxybenzotriazole and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.

Step 1: reacting a compound of general formula (I-F-M1A) with a compoundof general formula (KI3) in the presence of a condensing agent,optionally under a basic condition, to obtain a compound of generalformula (I-F-M2A)

Step 2: removing a protecting group of the compound of general formula(I-F-M2A) to obtain a compound of general formula (I-F-M3A)

Step 3: reacting a compound of general formula (KI1) with the compoundof general formula (I-F-M3A) in the presence of a condensing agent,optionally under a basic condition, to obtain the compound of generalformula (I-F)

wherein,

Re is an amino protecting group, preferably Fomc;

W, Y, Z, R^(L1), R^(L2), R¹, L¹, and L² are defined as in any formula(I-F) in embodiments of the sixth aspect.

Reagents that provide basic conditions include organic bases andinorganic bases, wherein the organic bases include, but are not limitedto, triethylamine, diethylamine, N-methylmorpholine, pyridine,piperidine, NA-diisopropylethylamine, n-butyllithium, lithiumdiisopropylamide, potassium acetate, sodium tert-butoxide, potassiumtert-butoxide, and the like, and the inorganic bases include, but arenot limited to, sodium hydride, potassium carbonate, sodium carbonate,cesium carbonate, sodium hydroxide, lithium hydroxide, and the like.

The condensing agent may be selected from the group consisting of4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride,1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride, N,N-dicyclohexylcarbodiimide,N,N′-diisopropylcarbodiimide, O-benzotriazol-N,N,N,N-tetramethyluroniumtetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazol,O-benzotriazol-N,N,N,N-tetramethyluronium hexafluorophosphate,2-(7-azobenzotriazol)-N,N,N,N-tetramethyluronium hexafluorophosphate,benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate,and benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate,preferably 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholiniumchloride, or 1-hydroxybenzotriazole and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.

wherein R¹ is selected from the group consisting of: —O—, —(R²)N—,—P(═O)(R²)— and —S—;

L² is —(C(R^(3a))(R^(3b)))_(m)—R,

wherein 0 or no less than 1 methylene unit of L² is independentlyreplaced by -Cy-, —N(R⁴)C(O)—, —C(O)N(R⁴)—, —C(O)—, —OC(O)—, —C(O)O—,—NR⁴—, —O—, —S—, —SO—, —SO₂—, —P(R⁴)—, —P(═O)(R⁴)—, —N(R⁴)SO₂—,—SO₂N(R⁴)—, —C(═S)—, —C(═NR⁴)—, —N═C— or —C(═N₂)—;

L¹ is —(C(R^(5a))(R^(5b)))_(n)—,

wherein 0 or no less than 1 methylene unit of L¹ is independentlyreplaced by -Cy-, —N(R⁶)C(O)—, —C(O)N(R⁶)—, —C(O)—, —OC(O)—, —C(O)O—,—NR⁶—, —O—, —S—, —SO—, —SO₂—, —P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—,—SO₂N(R⁶)—, —C(═S)—, —C(═NR⁶)—, —N═C— or —C(═N₂)—;

-Cy- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein -Cy-is unsubstituted or independently substituted with no less than 1substituent R⁷;

wherein each R^(3a), each R^(3b), each R⁴, each R^(5a), each R^(5b) andeach R⁶ are each independently hydrogen, protium, deuterium, tritium,halogen, —NO₂, —CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R,—C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic groupoptionally substituted with R; or, R^(3a) and R^(5a), R⁴ and R^(5a),R^(3a) and R⁶ or R⁴ and R⁶ each independently optionally form a ring Btogether with an atom therebetween, wherein the ring B is selected fromthe group consisting of: 5-8 membered heteroarylene and 3-10 memberedsaturated or partially unsaturated heterocyclylene, and the ring B isunsubstituted or independently substituted with no less than 1substituent R⁸;

wherein each R², each R⁷ and each R⁸ are each independently hydrogen,protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R,or a C₁₋₆ aliphatic group optionally substituted with R;

wherein each R, each R^(a) and each R^(b) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

m and n are each independently selected from the group consisting ofintegers ≥1.

Step 1: reacting a compound of general formula (I-F-M1B) with a compoundof general formula (KI3) in the presence of or in the absence of areducing agent, optionally under an acidic or basic condition, to obtaina compound of general formula (I-F-M2B)

Step 2: removing a protecting group of the compound of general formula(I-F-M2B) to obtain a compound of general formula (I-F-M3B)

Step 3: reacting a compound of general formula (KI1) with the compoundof general formula (I-F-M3B) in the presence of a condensing agent,optionally under a basic condition, to obtain the compound of generalformula (I-F)

wherein,

Re is an amino protecting group, preferably Fomc;

W, Y, Z, R^(L1), R^(L2), R¹, L¹, and L² are defined as in any formula(I-F) in embodiments of the sixth aspect.

Reducing agents include, but are not limited to, sodium hydride, calciumhydride, lithium hydride, lithium aluminum hydride, sodium borohydride,lithium borohydride, sodium triethylborohydride, sodiumtriacetoxyborohydride and sodium cyanoborohydride.

Reagents that provide acidic conditions include a protic acid and aLewis acid, wherein the protic acid includes, but is not limited to,hydrochloric acid, sulfuric acid, nitric acid, nitrous acid, sulfurousacid, phosphoric acid, phosphorous acid, formic acid, acetic acid,propionic acid, butyric acid, citric acid, benzoic acid,p-toluenesulfonic acid, p-nitrobenzoic acid, methanesulfonic acid,trifluoromethanesulfonic acid and trifluoroacetic acid, and the Lewisacid includes, but is not limited to, boron trifluoride, zinc chloride,magnesium chloride, aluminum chloride, stannic chloride and ferricchloride.

Reagents that provide basic conditions include organic bases andinorganic bases, wherein the organic bases include, but are not limitedto, triethylamine, diethylamine, N-methylmorpholine, pyridine,piperidine, NA-diisopropylethylamine, n-butyllithium, lithiumdiisopropylamide, potassium acetate, sodium tert-butoxide, potassiumtert-butoxide, and the like, and the inorganic bases include, but arenot limited to, sodium hydride, potassium carbonate, sodium carbonate,cesium carbonate, sodium hydroxide, lithium hydroxide, and the like.

The condensing agent may be selected from the group consisting of4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride,1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride, N,N-dicyclohexylcarbodiimide,N,N′-diisopropylcarbodiimide, O-benzotriazol-N,N,N,N-tetramethyluroniumtetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazol,O-benzotriazol-N,N,N,N-tetramethyluronium hexafluorophosphate,2-(7-azobenzotriazol)-N,N,N,N-tetramethyluronium hexafluorophosphate,benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate,and benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate,preferably 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholiniumchloride, or 1-hydroxybenzotriazole and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.

wherein R¹ is selected from the group consisting of: —O—, —(R²)N—,—P(═O)(R²)— and —S—;

L² is —(C(R^(3a))(R^(3b)))_(m)—R,

wherein 0 or no less than 1 methylene unit of L² is independentlyreplaced by -Cy-, —N(R⁴)C(O)—, —C(O)N(R⁴)—, —C(O)—, —OC(O)—, —C(O)O—,—NR⁴—, —O—, —S—, —SO—, —SO₂—, —P(R⁴)—, —P(═O)(R⁴)—, —N(R⁴)SO₂—,—SO₂N(R⁴)—, —C(═S)—, —C(═NR⁴)—, —N═C— or —C(═N₂)—;

L¹ is —(C(R^(5a))(R^(5b)))_(n)—,

wherein 0 or no less than 1 methylene unit of L¹ is independentlyreplaced by -Cy-, —N(R⁶)C(O)—, —C(O)N(R⁶)—, —C(O)—, —OC(O)—, —C(O)O—,—NR⁶—, —O—, —S—, —SO—, —SO₂—, —P(R⁶)—, —P(═O)(R⁶)—, —N(R⁶)SO₂—,—SO₂N(R⁶)—, —C(═S)—, —C(═NR⁶)—, —N═C— or —C(═N₂)—;

-Cy- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein -Cy-is unsubstituted or independently substituted with no less than 1substituent R⁷;

wherein each R^(3a), each R^(3b), each R⁴, each R^(5a), each R^(5b) andeach R⁶ are each independently hydrogen, protium, deuterium, tritium,halogen, —NO₂, —CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R,—C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)),—SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic groupoptionally substituted with R; or, R^(3a) and R^(5a), R⁴ and R^(5a),R^(3a) and R⁶ or R⁴ and R⁶ each independently optionally form a ring Btogether with an atom therebetween, wherein the ring B is selected fromthe group consisting of: 5-8 membered heteroarylene and 3-10 memberedsaturated or partially unsaturated heterocyclylene, and the ring B isunsubstituted or independently substituted with no less than 1substituent R⁸;

wherein each R², each R⁷ and each R⁸ are each independently hydrogen,protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(R^(b)), —OC(O)R, —N(R)SO₂R,or a C₁₋₆ aliphatic group optionally substituted with R;

wherein each R, each R^(a) and each R^(b) are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH,—NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H,—C(O)NH₂, —SO₂NH₂, —OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

m and n are each independently selected from the group consisting ofintegers ≥1.

Step 1: reacting a compound of general formula (I-E) with a compound ofgeneral formula (KI2), optionally under an acidic condition, to obtain acompound of general formula (I-F-M1C)

Step 2: removing a protecting group of the compound of general formula(I-F-M1C) to obtain a compound of general formula (I-F-M2C)

Step 3: reacting a compound of general formula (KI1) with the compoundof general formula (I-F-M2C) in the presence of a condensing agent,optionally under a basic condition, to obtain the compound of generalformula (I-F)

Re is an amino protecting group, preferably Fomc;

W, Y, Z, R^(L1), R^(L2), R¹, L¹, and L² are defined as in any generalformula (I-F) in embodiments of the sixth aspect.

Reducing agents include, but are not limited to, sodium hydride, calciumhydride, lithium hydride, lithium aluminum hydride, sodium borohydride,lithium borohydride, sodium triethylborohydride, sodiumtriacetoxyborohydride and sodium cyanoborohydride.

Reagents that provide acidic conditions include a protic acid and aLewis acid, wherein the protic acid includes, but is not limited to,hydrochloric acid, sulfuric acid, nitric acid, nitrous acid, sulfurousacid, phosphoric acid, phosphorous acid, formic acid, acetic acid,propionic acid, butyric acid, citric acid, benzoic acid,p-toluenesulfonic acid, p-nitrobenzoic acid, methanesulfonic acid,trifluoromethanesulfonic acid and trifluoroacetic acid, and the Lewisacid includes, but is not limited to, boron trifluoride, zinc chloride,magnesium chloride, aluminum chloride, stannic chloride and ferricchloride.

Reagents that provide basic conditions include organic bases andinorganic bases, wherein the organic bases include, but are not limitedto, triethylamine, diethylamine, N-methylmorpholine, pyridine,piperidine, NA-diisopropylethylamine, n-butyllithium, lithiumdiisopropylamide, potassium acetate, sodium tert-butoxide, potassiumtert-butoxide, and the like, and the inorganic bases include, but arenot limited to, sodium hydride, potassium carbonate, sodium carbonate,cesium carbonate, sodium hydroxide, lithium hydroxide, and the like.

The condensing agent may be selected from the group consisting of4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride,1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride, N,N-dicyclohexylcarbodiimide,N,N′-diisopropylcarbodiimide, O-benzotriazol-N,N,N,N-tetramethyluroniumtetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazol,O-benzotriazol-N,N,N,N-tetramethyluronium hexafluorophosphate,2-(7-azobenzotriazol)-N,N,N,N-tetramethyluronium hexafluorophosphate,benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate,and benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate,preferably 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholiniumchloride, or 1-hydroxybenzotriazole and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.

Step 1: reacting a compound of general formula (II-F-M1x) or a compoundof general formula (II-F-M1y) with a compound of general formula (KI4)in the presence of a condensing agent, optionally under a basiccondition, to obtain a compound of general formula (II-F-M2x) or generalformula (II-F-M2y)

Step 2: removing a protecting group of the compound of general formula(II-F-M2x) or general formula (II-F-M2y) to obtain a compound of generalformula (II-F-M3x) or general formula (II-F-M3y)

Step 3: reacting a compound of general formula (KI1) with the compoundof general formula (II-F-M3x) or general formula (II-F-M3y) in thepresence of a condensing agent, optionally under a basic condition, toobtain the compound of general formula (II-Fx) or general formula(II-Fy)

wherein,

Re is an amino protecting group, preferably Fomc;

W, Y, Z, R^(L1), R^(L2), A, X¹, L¹, L² and p are defined as in anyformula (II-Fx) in embodiments of the sixth aspect, or W, Y, Z, R^(L1),R^(L2), A, X¹, L¹, X² and q are defined as in any formula (II-Fy) inembodiments of the sixth aspect.

Reagents that provide basic conditions include organic bases andinorganic bases, wherein the organic bases include, but are not limitedto, triethylamine, diethylamine, N-methylmorpholine, pyridine,piperidine, NA-diisopropylethylamine, n-butyllithium, lithiumdiisopropylamide, potassium acetate, sodium tert-butoxide, potassiumtert-butoxide, and the like, and the inorganic bases include, but arenot limited to, sodium hydride, potassium carbonate, sodium carbonate,cesium carbonate, sodium hydroxide, lithium hydroxide, and the like.

The condensing agent may be selected from the group consisting of4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride,1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride, N,N-dicyclohexylcarbodiimide,N,N′-diisopropylcarbodiimide, O-benzotriazol-N,N,N,N-tetramethyluroniumtetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazol,O-benzotriazol-N,N,N,N-tetramethyluronium hexafluorophosphate,2-(7-azobenzotriazol)-N,N,N,N-tetramethyluronium hexafluorophosphate,benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate,and benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate,preferably 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholiniumchloride, or 1-hydroxybenzotriazole and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.

Step 1: reacting a compound of general formula (III-F-M1) with acompound of general formula (KI4) in the presence of a condensing agent,optionally under a basic condition, to obtain a compound of generalformula (III-F-M2)

Step 2: removing a protecting group of the compound of general formula(III-F-M2) to obtain a compound of general formula (III-F-M3)

Step 3: reacting a compound of general formula (KI1) with the compoundof general formula (III-F-M3) in the presence of a condensing agent,optionally under a basic condition, to obtain the compound of generalformula (III-F)

wherein,

Re is an amino protecting group, preferably Fomc;

W, Y, Z, R^(L1), R^(L2), R¹ and X are defined as in any general formula(III-F) in embodiments of the sixth aspect.

Reagents that provide basic conditions include organic bases andinorganic bases, wherein the organic bases include, but are not limitedto, triethylamine, diethylamine, N-methylmorpholine, pyridine,piperidine, NA-diisopropylethylamine, n-butyllithium, lithiumdiisopropylamide, potassium acetate, sodium tert-butoxide, potassiumtert-butoxide, and the like, and the inorganic bases include, but arenot limited to, sodium hydride, potassium carbonate, sodium carbonate,cesium carbonate, sodium hydroxide, lithium hydroxide, and the like.

The condensing agent may be selected from the group consisting of4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride,1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride, N,N-dicyclohexylcarbodiimide,N,N′-diisopropylcarbodiimide, O-benzotriazol-N,N,N,N-tetramethyluroniumtetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazol,O-benzotriazol-N,N,N,N-tetramethyluronium hexafluorophosphate,2-(7-azobenzotriazol)-N,N,N,N-tetramethyluronium hexafluorophosphate,benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate,and benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate,preferably 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholiniumchloride, or 1-hydroxybenzotriazole and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.

wherein,

Ab is a ligand, and after being reduced, Ab reacts with the generalformula (I-F) to obtain the compound of general formula (I-D);

reducing agents include, but are not limited to,tris(2-carboxyethyl)phosphine, mercaptoethanol, dithiothreitol,cysteine, reduced glutathione, and the like; in particular, disulfidebonds on the antibody are preferably reduced;

W, Y, Z, R^(L1), R^(L2), R¹, L¹ and L² are defined as in any formula(I-D) in embodiments of the fourth aspect.

wherein,

Ab is a ligand, and after being reduced, Ab reacts with the generalformula (II-F), including general formula (II-Fx) or general formula(II-Fy), to obtain the compound of general formula (II-Dx) or generalformula (II-Dy);

reducing agents include, but are not limited to,tris(2-carboxyethyl)phosphine, mercaptoethanol, dithiothreitol,cysteine, reduced glutathione, and the like; in particular, disulfidebonds on the antibody are preferably reduced;

W, Y, Z, R^(L1), R^(L2), A, X¹, L¹, L² and p a are defined as in anyformula (II-Dx) in embodiments of the fourth aspect, or W, Y, Z, R^(L1),R^(L2), A, X¹, L¹, X² and q are defined as in any formula (II-Dy) inembodiments of the fourth aspect.

wherein,

Ab is a ligand, and after being reduced, Ab reacts with the generalformula (III-F) to obtain the compound of general formula (III-D);

reducing agents include, but are not limited to,tris(2-carboxyethyl)phosphine, mercaptoethanol, dithiothreitol,cysteine, reduced glutathione, and the like; in particular, disulfidebonds on the antibody are preferably reduced;

W, Y, Z, R^(L1), R^(L2), R¹ and X are defined as in any general formula(III-D) in embodiments of the fourth aspect.

Technical Schemes

1. A compound or a tautomer, a mesomer, a racemate, an enantiomer or adiastereoisomer thereof, or a mixture thereof, or a pharmaceuticallyacceptable salt thereof, wherein the compound comprises a structureshown as formula (I-A):

wherein, R1 is selected from the group consisting of: —O—, —(R2)N—,—P(═O)(R2)-, —P(R2)- and —S—;

L² is —(C(R3a)(R3b))m-R,

wherein 0 or no less than 1 methylene unit of L² is independentlyreplaced by -Cy-, —N(R4)C(O)—, —C(O)N(R4)-, —C(O)—, —OC(O)—, —C(O)O—,—NR4-, —O—, —S—, —SO—, —SO₂—, —P(R4)-, —P(═O)(R4)-, —N(R4)SO₂—,—SO₂N(R4)-, —C(═S)—, —C(═NR4)-, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

L¹ is —(C(R5a)(R5b))n-,

wherein 0 or no less than 1 methylene unit of L¹ is independentlyreplaced by -Cy-, —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—, —C(O)O—,—NR6-, —O—, —S—, —SO—, —SO₂—, —P(R6)-, —P(═O)(R6)-, —N(R6)SO₂—,—SO₂N(R6)-, —C(═S)—, —C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N₂)—;

-Cy- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein -Cy-is unsubstituted or independently substituted with no less than 1substituent R7;

wherein each R3a, each R3b, each R4, each R5a, each R5b and each R6 areeach independently hydrogen, protium, deuterium, tritium, halogen, —NO₂,—CN, —OR, —SR, —N(R^(a))(R^(b)), —C(O)R, —CO₂R, —C(O)C(O)R,—C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(R^(a))(R^(b)), —SO₂N(R^(a))(Rb),—OC(O)R, —N(R)SO₂R, or a C₁₋₆ aliphatic group optionally substitutedwith R; or, R3a and R5a, R4 and R5a, R3a and R6 or R4 and R6 eachindependently optionally form a ring B together with an atomtherebetween, wherein the ring B is selected from the group consistingof: 5-8 membered heteroarylene and 3-10 membered saturated or partiallyunsaturated heterocyclylene, and the ring B is unsubstituted orindependently substituted with no less than 1 substituent R8;

wherein each R2, each R7 and each R8 are each independently hydrogen,protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(R^(a))(Rb), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R,—S(O)₂R, —C(O)N(R^(a))(Rb), —SO₂N(R^(a))(Rb), —OC(O)R, —N(R)SO₂R, or aC₁₋₆ aliphatic group optionally substituted with R;

wherein each R, each Ra and each Rb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH, —NH₂, —C(O)H,—CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H, —C(O)NH₂, —SO₂NH₂,—OC(O)H, —N(H)SO₂H or a C₁₋₆ aliphatic group;

m and n are each independently selected from the group consisting ofintegers ≥1.

2. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 1, wherein each R3a, each R3b, each R4, each R5a, each R5b andeach R6 are each independently hydrogen, protium, deuterium, tritium,halogen, —NO₂, —CN, —OR, —SR, —N(Ra)(Rb), —C(O)R, —CO₂R, —C(O)C(O)R,—C(O)CH₂C(O)R, —S(O)R, —S(O)₂R, —C(O)N(Ra)(Rb), —SO₂N(Ra)(Rb), —OC(O)R,—N(R)SO₂R, or a C₁₋₆ aliphatic group optionally substituted with R.

3. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 1, wherein R3a and R5a, R4 and R5a, R3a and R6, or R4 and R6 eachindependently optionally form a ring B together with an atomtherebetween, wherein the ring B is selected from the group consistingof: 5-8 membered heteroarylene and 3-10 membered saturated or partiallyunsaturated heterocyclylene, and the ring B is unsubstituted orsubstituted with no less than 1 substituent R8.

4. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-3, wherein m is 1 or 2.

5. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-4, wherein m is 1, and L² is —C(R3a)(R3b)-R.

6. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-5, wherein L² is —C(R3a)(R3b)-R, and 0 methyleneunits of L² are replaced by -Cy-, —N(R4)C(O)—, —C(O)N(R4)-, —C(O)—,—OC(O)—, —C(O)O—, —NR4-, —O—, —S—, —SO—, —SO2-, —P(R4)-, —P(═O)(R4)-,—N(R4)SO2-, —SO2N(R4)-, —C(═S)—, —C(═NR4)-, —N═N—, —C═N—, —N═C— or—C(═N₂)—.

7. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-4, wherein m is 2, and L2 is —(C(R3a)(R3b))2-R.

8. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1˜4 and 7, wherein L2 is —(C(R3a)(R3b))2-R, and 0methylene units of L² are replaced by -Cy-, —N(R4)C(O)—, —C(O)N(R4)-,—C(O)—, —OC(O)—, —C(O)O—, —NR4-, —O—, —S—, —SO—, —SO2-, —P(R4)-,—P(═O)(R4)-, —N(R4)SO2-, —SO₂N(R4)-, —C(═S)—, —C(═NR4)-, —N═N—, —C═N—,—N═C— or —C(═N2)-.

9. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-4 and 7, wherein L2 is —(C(R3a)(R3b))2-R, and 1methylene unit of L² is replaced by -Cy-, —N(R4)C(O)—, —C(O)N(R4)-,—C(O)—, —OC(O)—, —C(O)O—, —NR4-, —O—, —S—, —SO—, —SO2-, —P(R4)-,—P(═O)(R4)-, —N(R4)SO2-, —SO₂N(R4)-, —C(═S)—, —C(═NR4)-, —N═N—, —C═N—,—N═C— or —C(═N₂)—.

10. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-4, 7 and 9, wherein L2 is —(C(R3a)(R3b))₂-R, and 1methylene unit of L2 is replaced by —C(O)— or -Cy-.

11. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-4,7 and 9-10, wherein L2 is —(C(R3a)(R3b))₂-R, and 1methylene unit of L2 is replaced by —C(O)—.

12. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-4,7 and 9-11, wherein L2 is —C(O)—C(R3a)(R3b)-R.

13. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-4,7 and 9-10, wherein L2 is —(C(R3a)(R3b))₂-R, and 1methylene unit of L2 is replaced by -Cy-.

14. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-4,7, 9-10 and 13, wherein L2 is —C(R3a)(R3b)-Cy-R.

15. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 1, wherein 0 methylene units of L2 are replaced by -Cy-,—N(R4)C(O)—, —C(O)N(R4)-, —C(O)—, —OC(O)—, —C(O)O—, —NR4-, —O—, —S—,—SO—, —SO2-, —P(R4)-, —P(═O)(R4)-, —N(R4)SO2-, —SO2N(R4)-, —C(═S)—,—C(═NR4)-, —N═N—, —C═N—, —N═C— or —C(═N2)-.

16. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 1, wherein 1 methylene unit of L2 is replaced by -Cy-,—N(R4)C(O)—, —C(O)N(R4)-, —C(O)—, —OC(O)—, —C(O)O—, —NR4-, —O—, —S—,—SO—, —SO2-, —P(R4)-, —P(═O)(R4)-, —N(R4)SO2-, —SO2N(R4)-, —C(═S)—,—C(═NR4)-, —N═N—, —C═N—, —N═C— or —C(═N2)-.

17. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1 and 16, wherein 1 methylene unit of L2 is replacedby —C(O)— or -Cy-.

18. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1 and 16-17, wherein 1 methylene unit of L2 isreplaced by —C(O)—.

19. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-18, wherein n is 2,3 or 5.

20. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19, wherein n is 2, and L1 is —(C(R5a)(R5b))2-.

21. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-20, wherein L1 is —(C(R5a)(R5b))2-, and 0 methyleneunits of L¹ are replaced by -Cy-, —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—,—OC(O)—, —C(O)O—, —NR6-, —O—, —S—, —SO—, —SO2-, —P(R6)-, —P(═O)(R6)-,—N(R6)SO2-, —SO2N(R6)-, —C(═S)—, —C(═NR6)-, —N═N—, —C═N—, —N═C— or—C(═N2)-.

22. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19, wherein L1 is —(C(R5a)(R5b))2-, and 1 methyleneunit of L¹ is replaced by -Cy-, —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—,—OC(O)—, —C(O)O—, —NR6-, —O—, —S—, —SO—, —SO2-, —P(R6)-, —P(═O)(R6)-,—N(R6)SO2-, —SO2N(R6)-, —C(═S)—, —C(═NR6)-, —N═N—, —C═N—, —N═C— or—C(═N2)-.

23. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19 and 22, wherein L1 is —(C(R5a)(R5b))2-, and 1methylene unit of L¹ is replaced by —C(O)— or —C(═S)—.

24. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19 and 22-23, wherein L1 is —(C(R5a)(R5b))2-, and 1methylene unit of L1 is replaced by —C(O)—.

25. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19 and 22-24, wherein L1 is —C(R5a)(R5b)—C(O)—.

26. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19 and 22-23, wherein L1 is —(C(R5a)(R5b))2-, and 1methylene unit of L1 is replaced by —C(═S)—.

27. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19, 22-23 and 26, wherein L1 is —C(R5a)(R5b)—C(═S)—.

28. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19, wherein n is 3, and L1 is —(C(R5a)(R5b))3-.

29. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19 and 28, wherein L1 is —(C(R5a)(R5b))3-, and 0methylene units of L¹ are replaced by -Cy-, —N(R6)C(O)—, —C(O)N(R6)-,—C(O)—, —OC(O)—, —C(O)O—, —NR6-, —O—, —S—, —SO—, —SO2-, —P(R6)-,—P(═O)(R6)-, —N(R6)SO2-, —SO2N(R6)-, —C(═S)—, —C(═NR6)-, —N═N—, —C═N—,—N═C— or —C(═N2)-.

30. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19 and 28, wherein L1 is —(C(R5a)(R5b))3-, and 1methylene unit of L¹ is replaced by -Cy-, —N(R6)C(O)—, —C(O)N(R6)-,—C(O)—, —OC(O)—, —C(O)O—, —NR6-, —O—, —S—, —SO—, —SO2-, —P(R6)-,—P(═O)(R6)-, —N(R6)SO2-, —SO2N(R6)-, —C(═S)—, —C(═NR6)-, —N═N—, —C═N—,—N═C— or —C(═N2)-.

31. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19, 28 and 30, wherein L1 is —(C(R5a)(R5b))3-, and 1methylene unit of L¹ is replaced by —C(O)—.

32. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19, 28 and 30-31, wherein L1 is—(C(R5a)(R5b))2-C(O)—.

33. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19, wherein n is 5, and L1 is —(C(R5a)(R5b))5-.

34. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19 and 33, wherein L1 is —(C(R5a)(R5b))5-, and 1methylene unit of L¹ is replaced by -NR6- or —O—.

35. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19 and 33-34, wherein L1 is —(C(R5a)(R5b))5-, and 1methylene unit of L1 is replaced by -NR6-.

36. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19 and 33-35, wherein L1 is—(C(R5a)(R5b))2-NR6-(C(R5a)(R5b))2-.

37. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19 and 33-34, wherein L1 is —(C(R5a)(R5b))5-, and 1methylene unit of L1 is replaced by —O—.

38. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19, 33-34 and 37, wherein L1 is—(C(R5a)(R5b))2-O—(C(R5a)(R5b))2-.

39. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19 and 33, wherein L1 is —(C(R5a)(R5b))5-, and 2methylene units of L¹ are each independently replaced by —C(O)—,-NR6- or—O—.

40. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19,33 and 39, wherein L1 is —(C(R5a)(R5b))5-, and 2methylene units of L¹ are each independently replaced by —C(O)— or-NR6-.

41. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19, 33 and 39-40, wherein L1 is—C(R5a)(R5b)—C(O)—NR6-(C(R5a)(R5b))2-.

42. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19, 33 and 39-41, wherein L1 is—(C(R5a)(R5b))2-NR6-C(O)—C(R5a)(R5b)-.

43. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19, 33 and 39, wherein L1 is —(C(R5a)(R5b))5-, and 2methylene units of L1 are each independently replaced by —C(O)— or —O—.

44. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19, 33, 39 and 43, wherein L1 is—(C(R5a)(R5b))2-O—C(R5a)(R5b)—C(O)—.

45. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19 and 33, wherein L1 is —(C(R5a)(R5b))5-, and 3methylene units of L¹ are each independently replaced by —C(O)— or-NR6-.

46. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-19, 33 and 45, wherein L1 is—(C(R5a)(R5b))2-NR6-C(O)—C(O)—.

47. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-18, wherein 0 methylene units of L1 are replaced by—Cy-, —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—, —C(O)O—, —NR6-, —O—,—S—, —SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-, —SO2N(R6)-, —C(═S)—,—C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-.

48. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-18, wherein 1 methylene unit of L1 is replaced by-Cy-, —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—, —C(O)O—, —NR6-, —O—,—S—, —SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-, —SO2N(R6)-, —C(═S)—,—C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-.

49. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-18 and 48, wherein 1 methylene unit of L1 isreplaced by —C(O)—, —C(═S)—, —NR6- or —O—.

50. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-18 and 48-49, wherein 1 methylene unit of L1 isreplaced by —C(O)— or —C(═S)—.

51. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-18 and 48-50, wherein 1 methylene unit of L1 isreplaced by —C(O)—.

52. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-18, wherein 2 methylene units of L¹ are eachindependently replaced by -Cy-, —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—,—OC(O)—, —C(O)O—, —NR6-, —O—, —S—, —SO—, —SO2-, —P(R6)-, —P(═O)(R6)-,—N(R6)SO2-, —SO2N(R6)-, —C(═S)—, —C(═NR6)-, —N═N—, —C═N—, —N═C— or—C(═N2)-.

53. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-18 and 52, wherein 2 methylene units of L1 are eachindependently replaced by —C(O)—, —NR6- or —O—.

54. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-18 and 52-53, wherein 2 methylene units of L1 areeach independently replaced by —C(O)— or -NR6-.

55. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-18, wherein 3 methylene units of L¹ are eachindependently replaced by -Cy-, —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—,—OC(O)—, —C(O)O—, —NR6-, —O—, —S—, —SO—, —SO2-, —P(R6)-, —P(═O)(R6)-,—N(R6)SO2-, —SO2N(R6)-, —C(═S)—, —C(═NR6)-, —N═N—, —C═N—, —N═C— or—C(═N2)-.

56. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-18 and 55, wherein 3 methylene units of L1 are eachindependently replaced by —C(O)— or -NR6-.

57. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1 and 3, wherein R3a and R5a independently optionallyform a ring B together with an atom therebetween.

58. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1, 3 and 57, wherein ring B is 3-10 membered saturatedor partially unsaturated heterocyclylene.

59. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1, 3 and 57-58, wherein ring B is 3-6 memberedsaturated or partially unsaturated heterocyclylene.

60. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1, 3 and 57-59, wherein ring B is 5 membered saturatedor partially unsaturated heterocyclylene.

61. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1, 3 and 57-60, wherein ring B is 5 membered saturatedheterocyclylene.

62. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1 and 3, wherein R4 and R5a independently optionallyform a ring B together with an atom therebetween.

63. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1, 3 and 62, wherein ring B is 3-10 membered saturatedor partially unsaturated heterocyclylene.

64. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1,3 and 62-63, wherein ring B is 6 membered saturatedor partially unsaturated heterocyclylene.

65. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1,3 and 62-64, wherein ring B is 6 membered saturatedheterocyclylene.

66. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 1, wherein L2 is —C(R3a)(R3b)-R or —(C(R3a)(R3b))2-R, and L1 is—(C(R5a)(R5b))2-, —(C(R5a)(R5b))3- or —(C(R5a)(R5b))5-.

67. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1 and 66, wherein L2 is —(C(R3a)(R3b))2-R, and L¹ is—(C(R5a)(R5b))2-.

68. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1 and 66-67, wherein L2 is —(C(R3a)(R3b))2-R, L¹ is—(C(R5a)(R5b))2-, and 1 methylene unit of L1 is replaced by —C(O)—,—C(═S)—, —NR6- or —O—.

69. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1 and 66-68, wherein L2 is —(C(R3a)(R3b))2-R, L1 is—(C(R5a)(R5b))2-, and 1 methylene unit of L1 is replaced by —C(O)—.

70. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1 and 66-69, wherein L2 is —(C(R3a)(R3b))2-R, and L1is —C(R5a)(R5b)—C(O)—.

71. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1 and 66-70, wherein R3a and R5a independentlyoptionally form a ring B together with an atom therebetween.

72. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1 and 66-71, wherein ring B is 3-10 membered saturatedor partially unsaturated heterocyclylene.

73. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1 and 66-72, wherein ring B is 5 membered saturatedheterocyclylene.

74. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1 and 66, wherein L2 is —C(R3a)(R3b)-R, and L1 is—(C(R5a)(R5b))2-.

75. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1, 66 and 74, wherein L2 is —C(R3a)(R3b)-R, L1 is—(C(R5a)(R5b))2-, and 1 methylene unit of L1 is replaced by —C(O)—,—C(═S)—, —NR6- or —O—.

76. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1,66 and 74-75, wherein L2 is —C(R3a)(R3b)-R, L1 is—(C(R5a)(R5b))2-, and 1 methylene unit of L1 is replaced by —C(O)—.

77. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1,66 and 74-76, wherein L2 is —C(R3a)(R3b)-R, and L1is —C(R5a)(R5b)—C(O)—.

78. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-77, wherein R1 is selected from the group consistingof: —O—, —(R2)N- and —S—.

79. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-78, wherein R1 is —O—.

80. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-78, wherein R1 is —(R2)N—.

81. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-78 and 80, wherein R2 is hydrogen or a C₁₋₆aliphatic group.

82. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-78 and 80-81, wherein R1 is —HN—.

83. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-82, wherein -Cy- is 6-10 membered arylene.

84. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-83, wherein -Cy- is phenylene.

85. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-84, wherein R7 is hydrogen.

86. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 1, wherein R3a and R3b are independently hydrogen, or R3a and R5aindependently optionally form a ring B together with an atomtherebetween.

87. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1 and 86, wherein ring B is 3-10 membered saturated orpartially unsaturated heterocyclylene.

88. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1 and 86-87, wherein ring B is 5 membered saturatedheterocyclylene.

89. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 1, wherein R4 is hydrogen, or R4 and R5a independently optionallyform a ring B together with an atom therebetween.

90. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 1, wherein R4 is hydrogen, or R4 and R5a independently optionallyform a ring B together with an atom therebetween.

91. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1 and 86-90, wherein R8 is hydrogen.

92. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 1, wherein R, Ra and Rb are each independently hydrogen.

93. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 1, wherein the ligand-drug conjugate comprises the followinggroup of structures:

94. A compound or a tautomer, a mesomer, a racemate, an enantiomer or adiastereoisomer thereof, or a mixture thereof, or a pharmaceuticallyacceptable salt thereof, wherein the compound comprises a structureshown as formula (II-A):

wherein, X1 is selected from the group consisting of: N, P, andsaturated or unsaturated C; when X1 is saturated C, X1 is substitutedwith R^(n);

when X1 is saturated C, ring A is selected from the group consisting of:3-10 membered saturated or partially unsaturated heterocyclyl, and 3-10membered saturated or partially unsaturated carbocyclyl, wherein ring Ais substituted with 0 or no less than 1 substituent R1a;

or, when X1 is unsaturated C, ring A is selected from the groupconsisting of: 6-10 membered aryl, 5-8 membered heteroaryl, 3-10membered partially unsaturated heterocyclyl, and 3-10 membered partiallyunsaturated carbocyclyl, wherein ring A is substituted with 0 or no lessthan 1 substituent Rib;

or, when X1 is N or P, ring A is selected from the group consisting of:5-8 membered heteroaryl and 3-10 membered saturated or partiallyunsaturated heterocyclyl, wherein ring A is substituted with 0 or noless than 1 substituent R1c;

when ring A is selected from the group consisting of: 6-10 memberedaryl, 5-8 membered heteroaryl, and 3-10 membered saturated or partiallyunsaturated carbocyclyl, ring A is substituted with p L2, wherein L2 isnot R^(n);

or, when ring A is 3-10 membered saturated or partially unsaturatedheterocyclyl, ring A is substituted with p L2, or ring A comprises qring-forming heteroatom X2, and X2 is used for direct or indirectlinking of a ligand;

X2 is selected from the group consisting of: N and P;

L2 is -R2-L3-, and R2 is used for direct or indirect linking of aligand;

L3 is —(C(R3a)(R3b))m-, wherein when L3 comprises a methylene unit, 0 orno less than 1 methylene unit of L3 is independently replaced by—N(R4)C(O)—, —C(O)N(R4)-, —C(O)—, —OC(O)—, —C(O)O—, —NR4-, —O—, —S—,—SO—, —SO2-, —P(R4)-, —P(═O)(R4)-, —N(R4)SO2-, —SO2N(R4)-, —C(═S)—,—C(═NR4)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

R2 is selected from the group consisting of: —O—, —(R2a)N—, —S— and—P(═O)(R2a)-;

L1 is —(C(R5a)(R5b))n-, wherein when L1 comprises a methylene unit, 0 orno less than 1 methylene unit of L1 is independently replaced by—N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—, —C(O)O—, —NR6-, —O—, —S—,—SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-, —SO2N(R6)-, —C(═S)—,—C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

wherein each R1a, each R1b, each R1c, each R2a, each R3a, each R3b, eachR4, each R5a, each R5b, each R6 and each Rn are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR,—N(Ra)(Rb), —C(O)R, —CO2R, —C(O)C(O)R, —C(O)CH2C(O)R, —S(O)R, —S(O)2R,—C(O)N(Ra)(Rb), —SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or a C₁₋₆ aliphaticgroup optionally substituted with R;

wherein each R, each Ra and each Rb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH₂,—OC(O)H, —N(H)SO2H or a C₁₋₆ aliphatic group;

m and n are each independently selected from the group consisting ofintegers ≥0, and p and q are each independently selected from the groupconsisting of integers ≥1.

95. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 94, wherein X1 is saturated C.

96. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-95, wherein ring A is selected from the groupconsisting of: 3-10 membered saturated heterocyclyl and 3-10 memberedsaturated carbocyclyl.

97. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-96, wherein ring A is 3-10 membered saturatedcarbocyclyl.

98. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-97, wherein ring A is 3-6 membered saturatedcarbocyclyl.

99. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-98, wherein ring A is 4 membered saturatedcarbocyclyl.

100. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-98, wherein ring A is 6 membered saturatedcarbocyclyl.

101. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-96, wherein ring A is 3-10 membered saturatedheterocyclyl.

102. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-96 and 101, wherein ring A is 3-6 memberedsaturated heterocyclyl.

103. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-96 and 101-102, wherein ring A is 3 memberedsaturated heterocyclyl.

104. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-96 and 101-103, wherein ring A comprises 1heteroatom.

105. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-96 and 101-104, wherein ring A comprises 1 nitrogenatom.

106. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-96 and 101-102, wherein ring A is 5 memberedsaturated heterocyclyl.

107. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-96, 101-102 and 106, wherein ring A comprises 1heteroatom.

108. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-96, 101-102 and 106-107, wherein ring A comprises 1nitrogen atom.

109. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-108, wherein ring A is substituted with 0substituent R1a.

110. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 94, wherein X1 is unsaturated C.

111. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 110, wherein ring A is selected from the groupconsisting of: 6-10 membered aryl and 5-8 membered heteroaryl.

112. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 110-111, wherein ring A is 6-10 membered aryl.

113. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 110-112, wherein ring A is phenyl.

114. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 110-113, wherein ring A is substituted with 0substituent R1b.

115. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 94, wherein X1 is N or P.

116. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 94 or 115, wherein X1 is N.

117. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 115-116, wherein ring A is selected from thegroup consisting of: 5-8 membered heteroaryl and 3-10 membered saturatedheterocyclyl.

118. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 115-117, wherein ring A is 3-10 memberedsaturated heterocyclyl.

119. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 115-118, wherein ring A is 3-6 memberedsaturated heterocyclyl.

120. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 115-119, wherein ring A is 6 membered saturatedheterocyclyl.

121. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 115-120, wherein ring A independently comprises2 heteroatoms.

122. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 115-121, wherein ring A independently comprises2 nitrogen atoms.

123. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 115-121, wherein ring A is substituted with 0substituent R1c.

124. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 94, wherein the formula (II-A) is a structure shown as formula(II-Ax):

wherein, when ring A is selected from the group consisting of: 6-10membered aryl, 5-8 membered heteroaryl, and 3-10 membered saturated orpartially unsaturated carbocyclyl, ring A is substituted with p L2;

or, when ring A is 3-10 membered saturated or partially unsaturatedheterocyclyl, ring A is substituted with p L2.

125. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 94, wherein the formula (II-A) is a structure shown as formula(II-Ay):

wherein ring A is 3-10 membered saturated or partially unsaturatedheterocyclyl, ring A comprises q ring-forming heteroatom X2, and X2 isused for direct or indirect linking of a ligand.

126. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 94, wherein ring A is selected from the group consisting of: 6-10membered aryl, 5-8 membered heteroaryl, and 3-10 membered saturatedcarbocyclyl.

127. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 126, wherein ring A is selected from the groupconsisting of: phenyl and 3-6 membered saturated carbocyclyl.

128. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 126-127, wherein ring A is 3-6 memberedsaturated carbocyclyl.

129. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 126-128, wherein ring A is 4 membered saturatedcarbocyclyl or 6 membered saturated carbocyclyl.

130. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 126-127, wherein ring A is phenyl.

131. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 126-130, wherein ring A is substituted with noless than 1 L2.

132. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 126-131, wherein ring A is substituted with 1L2.

133. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 126, wherein ring A is 3-10 membered saturatedheterocyclyl.

134. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94,126 and 133, wherein ring A is 3-6 memberedsaturated heterocyclyl.

135. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94, 126 and 133-134, wherein ring A is 3 memberedsaturated heterocyclyl.

136. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94, 126 and 133-134, wherein ring A is 5 memberedsaturated heterocyclyl.

137. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94, 126 and 133-134, wherein ring A is 6 memberedsaturated heterocyclyl.

138. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 133-137, wherein ring A is substituted with no lessthan 1 L2.

139. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 133-138, wherein ring A is substituted with 1 L2.

140. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 94, wherein m is 0, 1 or 2.

141. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 140, wherein m is 0, and L3 is a covalent bond.

142. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 140, wherein m is 1, and L3 is —C(R5a)(R5b)-.

143. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94, 140 and 142, wherein 0 methylene units of L3 arereplaced by —N(R4)C(O)—, —C(O)N(R4)-, —C(O)—, —OC(O)—, —C(O)O—, —NR4-,—O—, —S—, —SO—, —SO2-, —P(R4)-, —P(═O)(R4)-, —N(R4)SO2-, —SO2N(R4)-,—C(═S)—, —C(═NR4)-, —N═N—, —C═N—, —N═C— or —C(═N2)-.

144. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 140, wherein m is 2, and L3 is—(C(R3a)(R3b))2-.

145. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94, 140 and 144, wherein 0 methylene units of L3 arereplaced by —N(R4)C(O)—, —C(O)N(R4)-, —C(O)—, —OC(O)—, —C(O)O—, —NR4-,—O—, —S—, —SO—, —SO2-, —P(R4)-, —P(═O)(R4)-, —N(R4)SO2-, —SO2N(R4)-,—C(═S)—, —C(═NR4)-, —N═N—, —C═N—, —N═C— or —C(═N2)-.

146. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 125, wherein ring A comprises no less than 1ring-forming heteroatom X2, and X2 is used for direct or indirectlinking of a ligand.

147. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94,125 and 146, wherein ring A comprises 1ring-forming heteroatom X2.

148. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94,125 and 146-147, wherein X2 is N.

149. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-148, wherein n is 0 or 1.

150. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-149, wherein n is 0, and L1 is a covalent bond.

151. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-149, wherein n is 1, and L1 is —C(R5a)(R5b)-.

152. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-149 and 151, wherein 1 methylene unit of L1 isreplaced by —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—, —C(O)O—, —NR6-,—O—, —S—, —SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-, —SO2N(R6)-,—C(═S)—, —C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-.

153. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-149 and 151-152, wherein 1 methylene unit of L1 isreplaced by —C(O)—.

154. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-153, wherein R2 is selected from the groupconsisting of: —O—, —(R2a)N- and —S—.

155. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-154, wherein R2 is —O—.

156. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-153, wherein R2 is —(R2A)N—.

157. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-153 and 156, wherein R2a is hydrogen.

158. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-153 and 156-157, wherein R2 is —HN—.

159. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94-109, wherein R1a is hydrogen.

160. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 110-114, wherein R1b is hydrogen.

161. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 94 and 115-123, wherein R1c is hydrogen.

162. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 94, wherein R3a and R3b are each independently hydrogen.

163. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 94, wherein R4 is hydrogen.

164. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 94, wherein R5a and R5b are each independently hydrogen.

165. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 94, wherein R6 is hydrogen.

166. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 94, wherein R, R^(a) and Rb are each independently hydrogen.

167. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 94, wherein the compound comprises the following group ofstructures:

168. A compound or a tautomer, a mesomer, a racemate, an enantiomer or adiastereoisomer thereof, or a mixture thereof, or a pharmaceuticallyacceptable salt thereof, wherein the compound comprises a structureshown as formula (III-A):

wherein R1 is selected from the group consisting of: —O—, —(R2)N—,—P(═O)(R2)- and —S—;

X is selected from the group consisting of: -L1-C(R1a)(R1b)—C(O)—,-L1-C(R1a)(R1b)—C(S)—, -L1-L0- and -L3-L2-;

L1 is —(C(R3a)(R3b))m-, wherein 0 or no less than 1 methylene unit of L1is independently replaced by —C(O)—, —C(═S)—, —C(═NR^(4b))- or —C(═N2)-;

L0 is —C(R2a)(R2b)-, or L0 is —C(═S)—, —C(═NR4a)- or —C(═N2)-;

L2 is —C(R5a)(R5b)-, wherein 0 or 1 methylene unit of L2 is replaced by—N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—, —C(O)O—, —NR6-, —O—, —S—,—SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-, —SO2N(R6)-, —C(═S)—,—C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

L3 is —(C(R7a)(R7b))n-, wherein no less than 1 methylene unit of L3 isindependently replaced by —N(R8)C(O)—, —C(O)N(R8)-, —OC(O)—, —C(O)O—,—NRB—, —O—, —S—, —SO—, —SO2-, —P(R8)-, —P(═O)(R8)-, —N(R8)SO2-,—SO2N(R8)-, —N═N—, —C═N— or —N═C—, and 0 or no less than 1 methyleneunit of L3 is also independently replaced by —C(O)—, —C(═S)—, —C(═NR8)-or —C(═N2)-;

wherein each R1a, each Rib, each R2, each R2a, each R2b, each R3a, eachR3b, each R4a, each R4b, each R5a, each R5b, each R6, each R7a, each R7band each R8 are each independently hydrogen, protium, deuterium,tritium, halogen, —NO₂, —CN, —OR, —SR, —N(Ra)(Rb), —C(O)R, —CO₂R,—C(O)C(O)R, —C(O)CH2C(O)R, —S(O)R, —S(O)2R, —C(O)N(Ra)(Rb),—SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or a C₁₋₆ aliphatic group optionallysubstituted with R;

wherein each R, each Ra and each Rb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH, —NH₂, —C(O)H,—CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H, —C(O)NH₂, —SO2NH₂,—OC(O)H, —N(H)SO2H or a C₁₋₆ aliphatic group;

m is selected from the group consisting of integers 0, and n is selectedfrom the group consisting of integers 1;

when R1 is —O— or —HN— and X is -L1-CH₂—C(O)—, no less than 1 methyleneunit of L1 is independently replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or—C(═N2)-, or each R3a and each R3b are not both hydrogen;

when R1 is —HN—, X is -L1-L0-, and L0 is —CH2-, no less than 1 methyleneunit of L1 is independently replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or—C(═N2)-, or each R3a and each R3b are not both hydrogen;

when R1 is —O—, X is -L3-C(O)—, and 1 methylene unit of L3 is replacedby —NRB, R8 is not —CH₂—CH₂—NH₂;

when R1 is —NH—, and X is -L3-C(O)—, no less than 1 methylene unit of L3is replaced by —N(R8)C(O)—, —OC(O)—, —C(O)O—, —S—, —SO—, —SO2-, —P(R8)-,—P(═O)(R8)-, —N(R8)SO2-, —SO2N(R8)-, —N═N—, —C═N— or —N═C—.

169. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein X is -L1-C(R1a)(R1b)—C(O)—, R1 is —S— or —(R2)N—,and R2 is not hydrogen.

170. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168-169, wherein L1 is —(C(R3a)(R3b))m-, and 0 or 1methylene unit of L1 is replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or—C(═N2)-.

171. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168-170, wherein 1 methylene unit of L1 is replaced by—C(O)—.

172. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein X is -L1-C(R1a)(R1b)—C(O)—, R1 is —O— or —HN—, m isnot 0, and no less than 1 methylene unit of L1 is replaced by —C(O)—,—C(═S)—, —C(═NR4b)- or —C(═N2)-.

173. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 172, wherein 1 methylene unit of L1 isreplaced by —C(O)—, —C(═S)—, —C(═NR4b)- or —C(═N2)-.

174. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 172-173, wherein 1 methylene unit of L1 isreplaced by —C(O)—.

175. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein X is -L1-C(R1a)(R1b)—C(O)—, R1 is —O— or —HN—, L1 is—(C(R3a)(R3b))m-, m is not 0,0 methylene units of L1 are replaced by—C(O)—, —C(═S)—, —C(═NR4b)- or —C(═N2)-, each R3a and each R3b are notboth hydrogen, and R1a and R1b are hydrogen.

176. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein X is -L1-C(R1a)(R1b)—C(O)—, and m is 0, 1 or 2.

177. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 176, wherein m is 0, and L1 is a covalentbond.

178. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 176-177, wherein R1 is (R2)N- or —S—, and R2is not hydrogen.

179. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 176-178, wherein R1 is —S—.

180. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 176, wherein m is 1, and L1 is —C(R3a)(R3b)-.

181. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 176 and 180, wherein 0 methylene units of L1 arereplaced by —C(O)—, —C(═S)—, —C(═NR4b)- or —C(═N2)-.

182. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 176 and 180-181, wherein R1 is (R2)N- or —S—, andR2 is not hydrogen.

183. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168,176 and 180-182, wherein R1 is —S—.

184. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168,176 and 180-183, wherein R1 is —S—, and R1a andR1b are hydrogen.

185. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168,176 and 180-183, wherein R1 is —S—, and R1a andR1b are each independently —N(R^(a))(Rb).

186. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168,176, 180-183 and 185, wherein R1a is —N(Ra)(Rb).

187. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168,176, 180-183 and 185-186, wherein R is hydrogen.

188. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168-187, wherein R1 is —(R2)N—, and R2 is a C1-6aliphatic group.

189. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168-188, wherein R2 is methyl.

190. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168-187, wherein L1 is —C(R3a)(R3b)-, R1 is —O— or—HN—, 0 methylene units of L1 are replaced by —C(O)—, —C(═S)—,—C(═NR4b)- or —C(═N2)-, and R3a and R3b are not both hydrogen.

191. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168-187 and 190, wherein R1 is —O—.

192. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168-187 and 190-191, wherein R3a is a C₁₋₆ aliphaticgroup.

193. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168-187 and 190-192, wherein R3a is a C₁₋₆ aliphaticgroup, and R3b is hydrogen or a C₁₋₆ aliphatic group.

194. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168-187 and 190-193, wherein R3a is a C₁₋₆ aliphaticgroup, and R3b is hydrogen.

195. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168-187 and 190-194, wherein R3a is methyl, and R3b ishydrogen.

196. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein R3a is a C₁₋₆ aliphatic group, and R3b is a C₁₋₆aliphatic group.

197. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 196, wherein R3a is methyl, and R3b is a C₁₋₆aliphatic group.

198. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 196-197, wherein R3a is methyl, and R3b ismethyl.

199. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 176, wherein m is 2, and L1 is—(C(R3a)(R3b))2-.

200. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 176 and 199, wherein 1 methylene unit of L1 isreplaced by —C(O)—, —C(═S)—, —C(═NR4b)- or —C(═N2)-.

201. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 176 and 199-200, wherein 1 methylene unit of L1is replaced by —C(O)—.

202. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 176 and 199-201, wherein R1 is selected from thegroup consisting of: —O—, —(R2)N—, —P(═O)(R2)- and —S—.

203. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 176 and 199-202, wherein R1 is —O—.

204. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 176 and 199-202, wherein R1 is —S—.

205. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 176 and 199-202, wherein R1 is —(R2)N—.

206. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 176, 199-202 and 205, wherein R2 is a C₁₋₆aliphatic group.

207. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 176, 199-202 and 205-206, wherein R2 is methyl.

208. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein X is -L1-L0-, R1 is —O—, —S— or —(R2)N—, and R2 isnot hydrogen.

209. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 208, wherein L1 is —C(R3a)(R3b), and 0 or 1methylene unit of L1 is replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or—C(═N2)-.

210. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 208-209, wherein 1 methylene unit of L1 isreplaced by —C(O)—.

211. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein X is -L1-L0-, R1 is —HN—, m is not 0, and L0 is—C(═S)—, —C(═NR4a)- or —C(═N2)-.

212. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein X is -L1-L0-, R1 is —HN—, m is not 0, L0 is—C(R2a)(R2b)-, L1 is —(C(R3a)(R3b))m-, and each R3a and each R3b are notboth hydrogen.

213. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein X is -L1-L0-, and m is 1 or 2.

214. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 213, wherein m is 1, and L1 is —C(R3a)(R3b)-.

215. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 213-214, wherein 0 methylene units of L1 arereplaced by —C(O)—, —C(═S)—, —C(═NR4b)- or —C(═N2)-.

216. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 213-215, wherein R1 is (R2)N- or —S—, and R2is not hydrogen.

217. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 213-216, wherein R1 is —S—.

218. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 213, wherein m is 2, and L1 is—(C(R3a)(R3b))2-.

219. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 213 and 218, wherein 0 methylene units of L1 arereplaced by —C(O)—, —C(═S)—, —C(═NR4b)- or —C(═N2)-.

220. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 213 and 218-219, wherein R1 is —O—, —S— or(R2)N—, and R2 is not hydrogen.

221. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 213 and 218-220, wherein R1 is —(R2)N—, and R2 isnot hydrogen.

222. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 213 and 218-221, wherein R2 is a C₁₋₆ aliphaticgroup.

223. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 213 and 218-222, wherein R2 is methyl.

224. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 213 and 218-220, wherein R1 is —O—.

225. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 213, 218-220 and 224, wherein R3a is a C₁₋₆aliphatic group.

226. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 213, 218-220 and 224-225, wherein R3a is a C₁₋₆aliphatic group, and R3b is hydrogen or a C₁₋₆ aliphatic group.

227. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 213, 218-220 and 224-226, wherein R3a is a C₁₋₆aliphatic group, and R3b is a C₁₋₆ aliphatic group.

228. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 213, 218-220 and 224-227, wherein R3a is methyl,and R3b is a C₁₋₆ aliphatic group.

229. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 213, 218-220 and 224-228, wherein R3a is methyl,and R3b is methyl.

230. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168, 213, 218-220 and 224-229, wherein L1 is—C(R3a)(R3b)—C(CH3)2-.

231. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein X is -L3-L2-, wherein L2 is —C(R5a)(R5b)-, L3 is—(C(R7a)(R7b))n-, R1 is —S— or —(R2)N—, and R2 is not hydrogen.

232. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein X is -L3-L2-, wherein L2 is —C(O)—, R1 is —O—, L3 is—(C(R7a)(R7b))n-, and when 1 methylene unit of L3 is replaced by —NRB,R8 is not a C₁₋₆ aliphatic group substituted with —NH₂.

233. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 232, wherein 1 methylene unit of L3 isreplaced by —NR8-, —O— or —SO—.

234. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 232-233, wherein R8 is a C₁₋₆ aliphatic group,and R8 is optionally substituted with hydrogen, protium, deuterium,tritium, halogen, —NO₂, —CN, —OH, —SH, —C(O)H, —CO₂H, —C(O)C(O)H,—C(O)CH₂C(O)H, —S(O)H, —S(O)₂H, —C(O)NH₂, —SO2NH₂, —OC(O)H, —N(H)SO2H ora C₁₋₆ aliphatic group.

235. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 232-234, wherein R8 is a C1-3 aliphatic group,and R8 is unsubstituted.

236. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein X is -L3-L2-, wherein when L2 is —C(O)—, R1 is —HN—,and L3 is —(C(R7a)(R7b))n-, at least 1 methylene unit of L3 is replacedby —N(R8)C(O)—, —OC(O)—, —C(O)O—, —S—, —SO—, —SO2-, —P(R8)-,—P(═O)(R8)-, —N(R8)SO2-, —SO2N(R8)-, —N═N—, —C═N— or —N═C—.

237. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein X is -L3-L2-, n is 4, and L3 is —(C(R7a)(R7b))4-.

238. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 237, wherein L2 is —C(R5a)(R5b)-, wherein 1methylene unit of L2 is replaced by —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—,—OC(O)—, —C(O)O—, —NR6, —O—, —S—, —SO—, —SO2-, —P(R6-, —P(═O)(R6-,—N(R6)SO2-, —SO2N(R6-, —C(═S)—, —C(═NR6-, —N═N—, —C═N—, —N═C— or—C(═N2)-.

239. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 237-238, wherein L2 is —C(R5a)(R5b)-, wherein1 methylene unit of L2 is replaced by —C(O)—, and L2 is —C(O)—.

240. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 237-239, wherein 1 methylene unit of L3 isreplaced by —N(R8)C(O)—, —C(O)N(R8)-, —OC(O)—, —C(O)O—, —NR8—, —O—, —S—,—SO—, —SO2-, —P(R8)-, —P(═O)(R8)-, —N(R8)SO2-, —SO2N(R8)-, —N═N—, —C═N—or —N═C—.

241. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 237-240, wherein 1 methylene unit of L3 isreplaced by —NR8—, —O— or —SO—.

242. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 237-241, wherein 1 methylene unit of L3 isreplaced by —NR8—.

243. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 237-242, wherein R8 is a C₁₋₆ aliphatic group.

244. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 237-243, wherein R8 is methyl.

245. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 237-244, wherein X is—(C(R7a)(R7b))2-N(CH₃)—C(R7a)(R7b)—C(O)—.

246. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 237-241, wherein 1 methylene unit of L3 isreplaced by —SO—.

247. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 237-238, wherein X is—(C(R7a)(R7b))2-SO—C(R7a)(R7b)—C(O)—.

248. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein 1 methylene unit of L3 is replaced by —O—.

249. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 248, wherein X is—(C(R7a)(R7b))2-O—C(R7a)(R7b)—C(O)—.

250. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein R1a and R1b are each independently hydrogen or—N(Ra)(Rb).

251. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein R2 is a C₁₋₆ aliphatic group.

252. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 251, wherein R2 is a C1-3 aliphatic group.

253. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 251-252, wherein R2 is methyl.

254. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein R3a and R3b are each independently hydrogen or aC₁₋₆ aliphatic group.

255. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 254, wherein R3a and R3b are eachindependently a C1-3 aliphatic group.

256. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 254-255, wherein R3a and R3b are methyl.

257. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein R4a and R4b are hydrogen.

258. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein R5a and R5b are hydrogen.

259. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein R6 is hydrogen.

260. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein R7a and R7b are hydrogen.

261. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein R8 is a C₁₋₆ aliphatic group.

262. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 261, wherein R8 is a C1-3 aliphatic group.

263. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 168 and 261-262, wherein R8 is methyl.

264. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein R, Ra and Rb are hydrogen.

265. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to technicalscheme 168, wherein the ligand-drug conjugate comprises the followinggroup of structures:

266. A compound or a tautomer, a mesomer, a racemate, an enantiomer or adiastereoisomer thereof, or a mixture thereof, or a pharmaceuticallyacceptable salt thereof, wherein the compound comprises a structureshown as formula (I-B):

wherein, Xa is nitrogen generated by removal of two hydrogen atoms froman amino group of a cytotoxic drug;

L is -La-Lb-Lc-;

-La- is selected from the group consisting of:

wherein W is —(C(Rwa)(Rwb))wn-, Y is —(OCH2CH2)yn-Oyp-, and Z is—(C(Rza)(Rzb))zn;

wherein wn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of W is independently replaced by-Cyr-, —N(Rwx)C(O)—, —C(O)N(Rwx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRwx-,—O—, —S—, —SO—, —SO2-, —P(Rwx)-, —P(═O)(Rwx)-, —N(Rwx)SO2-, —SO2N(Rwx)-,—C(═S)—, —C(═NRwx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

wherein yn is selected from the group consisting of integers 0, and ypis 0 or 1;

wherein zn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of Z is independently replaced by-Cyr-, —N(Rzx)C(O)—, —C(O)N(Rzx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRzx-,—O—, —S—, —SO—, —SO2-, —P(Rzx)-, —P(═O)(Rzx)-, —N(Rzx)SO2-, —SO2N(Rzx)-,—C(═S)—, —C(═NRzx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

-Cyr- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or independently substituted with no less than 1substituent Rcx;

wherein each Rwa, each Rwb, each Rza, each Rzb, each Rwx, each Rzx andeach Rcx are each independently hydrogen, protium, deuterium, tritium,halogen, —NO₂, —CN, —ORr, —SRr, —N(Rra)(Rrb), —C(O)Rr, —CO₂Rr,—C(O)C(O)Rr, —C(O)CH₂C(O)Rr, —S(O)Rr, —S(O)₂Rr, —C(O)N(Rra)(Rrb),—SO2N(Rra)(Rrb), —OC(O)Rr, —N(R)SO2Rr, or a C₁₋₆ aliphatic groupoptionally substituted with Rr;

wherein each Rr, each Rra and each Rrb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO₂, —CN, —OH, —SH, —NH₂, —C(O)H,—CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H, —S(O)₂H, —C(O)NH₂, —SO2NH₂,—OC(O)H, —N(H)SO2H or a C₁₋₆ aliphatic group;

-Lb- represents a peptide residue consisting of 2 to 7 amino acids;

-Lc- is selected from the group consisting of:

wherein RL1 and RL2 are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO₂,—CN, —OH, —SH, —NH₂, —C(O)H, —CO₂H, —C(O)C(O)H, —C(O)CH₂C(O)H, —S(O)H,—S(O)₂H, —C(O)NH₂, —SO2NH₂, —OC(O)H, —N(H)SO2H and a C₁₋₆ aliphaticgroup;

wherein R1 is selected from the group consisting of: —O—, —(R2)N—,—P(═O)(R2)- and —S—;

L2 is —(C(R3a)(R3b))m-R,

wherein 0 or no less than 1 methylene unit of L2 is independentlyreplaced by -Cy-, —N(R4)C(O)—, —C(O)N(R4)-, —C(O)—, —OC(O)—, —C(O)O—,—NR4-, —O—, —S—, —SO—, —SO2-, —P(R4)-, —

P(═O)(R4)-, —N(R4)SO2-, —SO2N(R4)-, —C(═S)—, —C(═NR4)-, —N═N—, —C═N—,—N═C— or —C(═N2)-;

L1 is —(C(R5a)(R5b))n-,

wherein 0 or no less than 1 methylene unit of L1 is independentlyreplaced by -Cy-, —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—, —C(O)O—,—NR6-, —O—, —S—, —SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-,—SO2N(R6)-, —C(═S)—, —C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

-Cy- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein -Cy-is unsubstituted or independently substituted with no less than 1substituent R7;

wherein each R3a, each R3b, each R4, each R5a, each R5b and each R6 areeach independently hydrogen, protium, deuterium, tritium, halogen, —NO₂,—CN, —OR, —SR, —N(Ra)(Rb), —C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R,—S(O)R, —S(O)₂R, —C(O)N(Ra)(Rb), —SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or aC₁₋₆ aliphatic group optionally substituted with R; or, R3a and R5a, R4and R5a, R3a and R6 or R4 and R6 each independently optionally form aring B together with an atom therebetween, wherein the ring B isselected from the group consisting of: 5-8 membered heteroarylene and3-10 membered saturated or partially unsaturated heterocyclylene, andthe ring B is unsubstituted or substituted with no less than 1substituent R8;

wherein each R2, each R7 and each R8 are each independently hydrogen,protium, deuterium, tritium, halogen, —NO₂, —CN, —OR, —SR, —N(Ra)(Rb),—C(O)R, —CO₂R, —C(O)C(O)R, —C(O)CH₂C(O)R, —S(O)R, —S(O)₂R,—C(O)N(Ra)(Rb), —SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or a C₁₋₆ aliphaticgroup optionally substituted with R;

wherein each R, each Ra and each Rb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

m and n are each independently selected from the group consisting ofintegers 1.

267. A compound or a tautomer, a mesomer, a racemate, an enantiomer or adiastereoisomer thereof, or a mixture thereof, or a pharmaceuticallyacceptable salt thereof, wherein the compound comprises a structureshown as formula (II-Bx) or formula (II-By):

wherein, Xa is nitrogen generated by removal of two hydrogen atoms froman amino group of a cytotoxic drug;

L is -La-Lb-Lc-;

-La- is selected from the group consisting of:

wherein W is —(C(Rwa)(Rwb))wn-, Y is —(OCH2CH₂)yn-Oyp-, and Z is—(C(Rza)(Rzb))zn;

wherein wn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of W is independently replaced by-Cyr-, —N(Rwx)C(O)—, —C(O)N(Rwx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRwx-,—O—, —S—, —SO—, —SO2-, —P(Rwx)-, —P(═O)(Rwx)-, —N(Rwx)SO2-, —SO2N(Rwx)-,—C(═S)—, —C(═NRwx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

wherein yn is selected from the group consisting of integers 0, and ypis 0 or 1;

wherein zn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of Z is independently replaced by-Cyr-, —N(Rzx)C(O)—, —C(O)N(Rzx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRzx-,—O—, —S—, —SO—, —SO2-, —P(Rzx)-, —P(═O)(Rzx)-, —N(Rzx)SO2-, —SO2N(Rzx)-,—C(═S)—, —C(═NRzx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

-Cyr- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or independently substituted with no less than 1substituent Rcx;

wherein each Rwa, each Rwb, each Rza, each Rzb, each Rwx, each Rzx andeach Rcx are each independently hydrogen, protium, deuterium, tritium,halogen, —NO2, —CN, —ORr, —SRr, —N(Rra)(Rrb), —C(O)Rr, —CO2Rr,—C(O)C(O)Rr, —C(O)CH2C(O)Rr, —S(O)Rr, —S(O)2Rr, —C(O)N(Rra)(Rrb),—SO2N(Rra)(Rrb), —OC(O)Rr, —N(R)SO2Rr, or a C1-6 aliphatic groupoptionally substituted with Rr;

wherein each Rr, each Rra and each Rrb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH₂, —SO2NH₂,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

-Lb- represents a peptide residue consisting of 2 to 7 amino acids;

-Lc- is selected from the group consisting of:

wherein RL1 and RL2 are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO2,—CN, —OH, —SH, —NH2, —C(O)H, —CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H,—S(O)2H, —C(O)NH2, —SO2NH2, —OC(O)H, —N(H)SO2H and a C1-6 aliphaticgroup;

wherein, X1 is selected from the group consisting of: N, P, andsaturated or unsaturated C; when X1 is saturated C, X1 is substitutedwith Rn;

when X1 is saturated C, ring A is selected from the group consisting of:3-10 membered saturated or partially unsaturated heterocyclyl, and 3-10membered saturated or partially unsaturated carbocyclyl, wherein ring Ais unsubstituted or substituted with no less than 1 substituent R1a;

or, when X1 is unsaturated C, ring A is selected from the groupconsisting of: 6-10 membered aryl, 5-8 membered heteroaryl, 3-10membered partially unsaturated heterocyclyl, and 3-10 membered partiallyunsaturated carbocyclyl, wherein ring A is unsubstituted or substitutedwith no less than 1 substituent R1b;

or, when X1 is N or P, ring A is selected from the group consisting of:5-8 membered heteroaryl and 3-10 membered saturated or partiallyunsaturated heterocyclyl, wherein ring A is unsubstituted or substitutedwith no less than 1 substituent R1c;

when ring A is selected from the group consisting of: 6-10 memberedaryl, 5-8 membered heteroaryl, and 3-10 membered saturated or partiallyunsaturated carbocyclyl, ring A is substituted with p L2, wherein L2 isnot R^(n);

or, when ring A is 3-10 membered saturated or partially unsaturatedheterocyclyl, ring A is substituted with p L2, or ring A comprises qring-forming heteroatom X2, and X2 is used for direct or indirectlinking of a ligand;

X2 is selected from the group consisting of: N and P;

L2 is -R2-L3-, and R2 is used for direct or indirect linking of aligand;

L3 is —(C(R3a)(R3b))m-, wherein 0 or no less than 1 methylene unit of L3is independently replaced by —N(R4)C(O)—, —C(O)N(R4)-, —C(O)—, —OC(O)—,—C(O)O—, —NR4-, —O—, —S—, —SO—, —SO2-, —P(R4)-, —P(═O)(R4)-, —N(R4)SO2-,—SO2N(R4)-, —C(═S)—, —C(═NR4)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

R2 is selected from the group consisting of: —O—, —(R2a)N—, —S— and—P(═O)(R2a)-;

L1 is —(C(R5a)(R5b))n —, wherein 0 or no less than 1 methylene unit ofL1 is independently replaced by —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—,—OC(O)—, —C(O)O—, —NR6-, —O—, —S—, —SO—, —SO2-, —P(R6)-, —P(═O)(R6)-,—N(R6)SO2-, —SO2N(R6)-, —C(═S)—, —C(═NR6)-, —N═N—, —C═N—, —N═C— or—C(═N2)-;

wherein each R1a, each R1b, each R1c, each R2a, each R3a, each R3b, eachR4, each R5a, each R5b, each R6 and each Rn are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO2, —CN, —OR, —SR,—N(Ra)(Rb), —C(O)R, —CO2R, —C(O)C(O)R, —C(O)CH2C(O)R, —S(O)R, —S(O)2R,—C(O)N(Ra)(Rb), —SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or a C1-6 aliphaticgroup optionally substituted with R;

wherein each R, each Ra and each Rb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

m and n are each independently selected from the group consisting ofintegers 0, and p and q are each independently selected from the groupconsisting of integers 1.

268. A compound or a tautomer, a mesomer, a racemate, an enantiomer or adiastereoisomer thereof, or a mixture thereof, or a pharmaceuticallyacceptable salt thereof, wherein the compound comprises a structureshown as formula (III-B):

wherein, Xa is nitrogen generated by removal of two hydrogen atoms froman amino group of a cytotoxic drug;

L is -La-Lb-Lc-;

-La- is selected from the group consisting of:

wherein W is —(C(Rwa)(Rwb))wn-, Y is —(OCH₂CH2)yn-Oyp, and Z is—(C(Rza)(Rzb))zn;

wherein wn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of W is independently replaced by-Cyr-, —N(Rwx)C(O)—, —C(O)N(Rwx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRwx-,—O—, —S—, —SO—, —SO2-, —P(Rwx)-, —P(═O)(Rwx)-, —N(Rwx)SO2-, —SO2N(Rwx)-,—C(═S)—, —C(═NRwx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

wherein yn is selected from the group consisting of integers 0, and ypis 0 or 1;

wherein zn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of Z is independently replaced by-Cyr-, —N(Rzx)C(O)—, —C(O)N(Rzx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRzx-,—O—, —S—, —SO—, —SO2-, —P(Rzx)-, —P(═O)(Rzx)-, —N(Rzx)SO2-, —SO2N(Rzx)-,—C(═S)—, —C(═NRzx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

-Cyr- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or independently substituted with no less than 1substituent Rcx;

wherein each Rwa, each Rwb, each Rza, each Rzb, each Rwx, each Rzx andeach Rcx are each independently hydrogen, protium, deuterium, tritium,halogen, —NO2, —CN, —ORr, —SRr, —N(Rra)(Rrb), —C(O)Rr, —CO2Rr,—C(O)C(O)Rr, —C(O)CH2C(O)Rr, —S(O)Rr, —S(O)2Rr, —C(O)N(Rra)(Rrb),—SO2N(Rra)(Rrb), —OC(O)Rr, —N(R)SO2Rr, or a C1-6 aliphatic groupoptionally substituted with Rr;

wherein each Rr, each Rra and each Rrb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

-Lb- represents a peptide residue consisting of 2 to 7 amino acids;

-Lc- is selected from the group consisting of:

wherein RL1 and RL2 are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO2,—CN, —OH, —SH, —NH2, —C(O)H, —CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H,—S(O)2H, —C(O)NH2, —SO2NH2, —OC(O)H, —N(H)SO2H and a C1-6 aliphaticgroup;

wherein R1 is selected from the group consisting of: —O—, —(R2)N—,—P(═O)(R2)- and —S—;

X is selected from the group consisting of: -L1-C(R1a)(R1b)—C(O)—,-L1-C(R1a)(R1b)—C(S)—, -L1-L0- and -L3-L2-;

L1 is —(C(R3a)(R3b))m-, wherein 0 or no less than 1 methylene unit of L1is independently replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or —C(═N2)-;

L0 is —C(R2a)(R2b)-, or L0 is —C(═S)—, —C(═NR4a)- or —C(═N2)-;

L2 is —C(R5a)(R5b)-, wherein 0 or 1 methylene unit of L2 is replaced by—N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—, —C(O)O—, —NR6-, —O—, —S—,—SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-, —SO2N(R6)-, —C(═S)—,—C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

L3 is —(C(R7a)(R7b))n-, wherein no less than 1 methylene unit of L3 isindependently replaced by —N(R8)C(O)—, —C(O)N(R8)-, —OC(O)—, —C(O)O—,—NRB—, —O—, —S—, —SO—, —SO2-, —P(R8)-, —P(═O)(R8)-, —N(R8)SO2-,—SO2N(R8)-, —N═N—, —C═N— or —N═C—, and 0 or no less than 1 methyleneunit of L3 is also independently replaced by —C(O)—, —C(═S)—, —C(═NR8)-or —C(═N2)-;

wherein each R1a, each R1b, each R2, each R2a, each R2b, each R3a, eachR3b, each R4a, each R4b, each R5a, each R5b, each R6, each R7a, each R7band each R8 are each independently hydrogen, protium, deuterium,tritium, halogen, —NO2, —CN, —OR, —SR, —N(R^(a))(Rb), —C(O)R, —CO2R,—C(O)C(O)R, —C(O)CH2C(O)R, —S(O)R, —S(O)2R, —C(O)N(R^(a))(Rb),—SO2N(R^(a))(Rb), —OC(O)R, —N(R)SO2R, or a C1-6 aliphatic groupoptionally substituted with R;

wherein each R, each Ra and each Rb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

m is selected from the group consisting of integers 0, and n is selectedfrom the group consisting of integers 1;

when R1 is —O— or —HN— and X is -L1-CH2-C(O)—, no less than 1 methyleneunit of L1 is independently replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or—C(═N2)-, or each R3a and each R3b are not both hydrogen;

when R1 is —HN—, X is -L1-L0-, and L0 is —CH2-, no less than 1 methyleneunit of L1 is independently replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or—C(═N2)-, or each R3a and each R3b are not both hydrogen;

when R1 is —O—, X is -L3-C(O)—, and 1 methylene unit of L3 is replacedby —NRB, R8 is not —CH2-CH2-NH2;

when R1 is —NH—, and X is -L3-C(O)—, no less than 1 methylene unit of L3is replaced by —N(R8)C(O)—, —OC(O)—, —C(O)O—, —S—, —SO—, —SO2-, —P(R8)-,—P(═O)(R8)-, —N(R8)SO2-, —SO2N(R8)-, —N═N—, —C═N— or —N═C—.

269. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 266-268, wherein

-La- is selected from the group consisting of:

wherein W is —(C(Rwa)(Rwb))wn-, Y is —(OCH2CH2)yn-Oyp-, and Z is—(C(Rza)(Rzb))zn;

wherein wn is selected from the group consisting of integers from 2 to6, and

0 or 1 methylene unit of W is independently replaced by -Cyr-,—N(Rwx)C(O)—, —C(O)N(Rwx)-, —C(O)—, —NRwx- or —O—;

wherein yn is selected from the group consisting of integers from 0 to12, and yp is 0 or 1;

wherein zn is selected from the group consisting of integers from 0 to10, and

0 or 1 methylene unit of Z is independently replaced by -Cyr-,—N(Rzx)C(O)—, —C(O)N(Rzx)—or —C(O)—;

-Cyr- is selected from the group consisting of: 6-10 membered aryleneand 3-10 membered saturated or partially unsaturated carbocyclylene,wherein -Cyr- is unsubstituted or independently substituted with 1 to 3substituent Rcx;

wherein each Rwa, each Rwb, each Rza, each Rzb, each Rwx, each Rzx andeach Rcx are each independently hydrogen, protium, deuterium, tritium,halogen, —NO2, —CN, —ORr, —SRr, —N(Rra)(Rrb), —C(O)Rr, —CO2Rr,—C(O)C(O)Rr, —C(O)CH2C(O)Rr, —S(O)Rr, —S(O)2Rr, —C(O)N(Rra)(Rrb),—SO2N(Rra)(Rrb), —OC(O)Rr, —N(R)SO2Rr, or a C1-6 aliphatic groupoptionally substituted with Rr;

wherein each Rr, each Rra and each Rrb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

-Lb- represents a peptide residue consisting of 2 to 7 amino acids, andthe peptide residue of -Lb- is a peptide residue formed of amino acidsselected from the group consisting of: phenylalanine, glycine, alanine,valine, citrulline, lysine, serine, glutamic acid and aspartic acid;

-Lc- is selected from the group consisting of:

wherein RL1 and RL2 are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO2,—CN, —OH, —SH, —NH2, —C(O)H, —CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H,—S(O)2H, —C(O)NH2, —SO2NH2, —OC(O)H, —N(H)SO2H and a C1-6 aliphaticgroup.

270. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 266-269, wherein

-La- is

wherein W is —(C(Rwa)(Rwb))wn-, Y is —(OCH2CH2)yn-Oyp-, and Z is—(C(Rza)(Rzb))zn;

wherein wn is 1, 2, 3 or 6, and

1 methylene unit of W is independently replaced by -Cyr-, —N(Rwx)C(O)—,—C(O)N(Rwx)- or —C(O)—;

wherein yn is 0, 4 or 8, and yp is 0 or 1;

wherein zn is 1, 2 or 3, and

1 methylene unit of Z is independently replaced by -Cyr-, —N(Rzx)C(O)—,—C(O)N(Rzx)- or —C(O)—;

-Cyr- is 3-10 membered saturated carbocyclylene, wherein -Cyr- isunsubstituted or independently substituted with 1 to 3 substituent Rcx;

wherein each Rwa, each Rwb, each Rza, each Rzb, each Rwx, each Rzx andeach Rcx are each independently hydrogen, halogen, —ORr, or a C1-6aliphatic group optionally substituted with Rr;

wherein each Rr is independently hydrogen, halogen or a C1-6 aliphaticgroup;

-Lb- represents a peptide residue consisting of 2 to 4 amino acids, andthe peptide residue of -Lb- is a peptide residue formed of amino acidsselected from the group consisting of: phenylalanine, glycine, alanine,valine, citrulline and lysine;

-Lc- is selected from the group consisting of:

wherein RL1 and RL2 are each independently selected from the groupconsisting of: hydrogen, halogen, —OH and a C1-6 aliphatic group.

271. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 266-270, wherein

-La- is

wherein W is —(C(Rwa)(Rwb))wn-, Y is —(OCH2CH2)yn-Oyp-, and Z is—(C(Rza)(Rzb))zn;

wherein wn is 1, 2, 3 or 6, and

1 methylene unit of W is independently replaced by -Cyr-, —N(Rwx)C(O)—,—C(O)N(Rwx)- or —C(O)—;

wherein yn is 0, 4 or 8, and yp is 0 or 1;

wherein zn is 1, 2 or 3, and

1 methylene unit of Z is independently replaced by -Cyr-, —N(Rzx)C(O)—,—C(O)N(Rzx)- or —C(O)—;

-Cyr- is 3-10 membered saturated carbocyclylene, wherein -Cyr- isunsubstituted or independently substituted with 1 to 3 substituent Rcx;

wherein each Rwa, each Rwb, each Rza, each Rzb, each Rwx, each Rzx andeach Rcx are each independently hydrogen, halogen, —ORr, or a C1-6aliphatic group optionally substituted with Rr;

wherein each Rr is independently hydrogen, halogen or a C1-6 aliphaticgroup;

-Lb- is selected from the group consisting of:

-La- is

wherein RL1 and RL2 are each independently selected from the groupconsisting of: hydrogen, halogen, —OH and a C1-6 aliphatic group.

272. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 266-271, wherein

-La- is

wherein W is —(C(Rwa)(Rwb))wn-, Y is —(OCH2CH2)yn-Oyp-, and Z is—(C(Rza)(Rzb))zn;

wherein wn is 1, 2, 3 or 6, and

1 methylene unit of W is independently replaced by -Cyr-, —N(Rwx)C(O)—,—C(O)N(Rwx)- or —C(O)—;

wherein yn is 0, 4 or 8, and yp is 0 or 1;

wherein zn is 1, 2 or 3, and

1 methylene unit of Z is independently replaced by -Cyr-, —N(Rzx)C(O)—,—C(O)N(Rzx)- or —C(O)—;

-Cyr- is 3-10 membered saturated carbocyclylene, wherein -Cyr- isunsubstituted or independently substituted with 1 to 3 substituent Rcx;

wherein each Rwa, each Rwb, each Rza, each Rzb, each Rwx, each Rzx andeach Rcx are each independently hydrogen, halogen, —ORr, or a C1-6aliphatic group optionally substituted with Rr;

wherein each Rr is independently hydrogen, halogen or a C1-6 aliphaticgroup;

-L_(b)- is

-L_(c)- is

wherein RL1 and RL2 are each independently selected from the groupconsisting of: hydrogen, halogen, —OH and a C1-6 aliphatic group.

273. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 266-272, wherein

-La- is

274. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 266-273, wherein

-Lb- is:

275. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 266-274, wherein

-Lc- is

276. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 266-275, wherein

-La-Lb-Lc- is

277. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 266-276, wherein the cytotoxic drug is shown asformula (EXA):

278. A compound or a tautomer, a mesomer, a racemate, an enantiomer or adiastereoisomer thereof, or a mixture thereof, or a pharmaceuticallyacceptable salt thereof, wherein the compound comprises a structureshown as formula (I-C):

wherein, L is -La-Lb-Lc-;

-La- is selected from the group consisting of:

wherein W is —(C(Rwa)(Rwb))wn-, Y is —(OCH2CH2)yn-Oyp-, and Z is—(C(Rza)(Rzb))zn;

wherein wn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of W is independently replaced by-Cyr-, —N(Rwx)C(O)—, —C(O)N(Rwx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRwx-,—O—, —S—, —SO—, —SO2-, —P(Rwx)-, —P(═O)(Rwx)-, —N(Rwx)SO2-, —SO2N(Rwx)-,—C(═S)—, —C(═NRwx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

wherein yn is selected from the group consisting of integers 0, and ypis 0 or 1;

wherein zn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of Z is independently replaced by-Cyr-, —N(Rzx)C(O)—, —C(O)N(Rzx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRzx-,—O—, —S—, —SO—, —SO2-, —P(Rzx)-, —P(═O)(Rzx)-, —N(Rzx)SO2-, —SO2N(Rzx)-,—C(═S)—, —C(═NRzx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

-Cyr- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or independently substituted with no less than 1substituent Rcx;

wherein each Rwa, each Rwb, each Rza, each Rzb, each Rwx, each Rzx andeach Rcx are each independently hydrogen, protium, deuterium, tritium,halogen, —NO2, —CN, —ORr, —SRr, —N(Rra)(Rrb), —C(O)Rr, —CO2Rr,—C(O)C(O)Rr, —C(O)CH2C(O)Rr, —S(O)Rr, —S(O)2Rr, —C(O)N(Rra)(Rrb),—SO2N(Rra)(Rrb), —OC(O)Rr, —N(R)SO2Rr, or a C1-6 aliphatic groupoptionally substituted with Rr;

wherein each Rr, each Rra and each Rrb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

-Lb- represents a peptide residue consisting of 2 to 7 amino acids;

-Lc- is selected from the group consisting of:

wherein RL1 and RL2 are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO2,—CN, —OH, —SH, —NH2, —C(O)H, —CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H,—S(O)2H, —C(O)NH2, —SO2NH2, —OC(O)H, —N(H)SO2H and a C1-6 aliphaticgroup;

wherein R1 is selected from the group consisting of: —O—, —(R2)N—,—P(═O)(R2)- and —S—;

L2 is —(C(R3a)(R3b))m-R,

wherein 0 or no less than 1 methylene unit of L2 is independentlyreplaced by -Cy-, —N(R4)C(O)—, —C(O)N(R4)-, —C(O)—, —OC(O)—, —C(O)O—,—NR4-, —O—, —S—, —SO—, —SO2-, —P(R4)-, —P(═O)(R4)-, —N(R4)SO2-,—SO2N(R4)-, —C(═S)—, —C(═NR4)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

L1 is —(C(R5a)(R5b))n-,

wherein 0 or no less than 1 methylene unit of L1 is independentlyreplaced by -Cy-, —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—, —C(O)O—,—NR6-, —O—, —S—, —SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-,—SO2N(R6)-, —C(═S)—, —C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

-Cy- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein -Cy-is unsubstituted or independently substituted with no less than 1substituent R7;

wherein each R3a, each R3b, each R4, each R5a, each R5b and each R6 areeach independently hydrogen, protium, deuterium, tritium, halogen, —NO2,—CN, —OR, —SR, —N(Ra)(Rb), —C(O)R, —CO2R, —C(O)C(O)R, —C(O)CH2C(O)R,—S(O)R, —S(O)2R, —C(O)N(Ra)(Rb), —SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or aC1-6 aliphatic group optionally substituted with R; or, R3a and R5a, R4and R5a, R3a and R6 or R4 and R6 each independently optionally form aring B together with an atom therebetween, wherein the ring B isselected from the group consisting of: 5-8 membered heteroarylene and3-10 membered saturated or partially unsaturated heterocyclylene, andthe ring B is unsubstituted or substituted with no less than 1substituent R8;

wherein each R2, each R7 and each R8 are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OR, —SR, —N(Ra)(Rb),—C(O)R, —CO2R, —C(O)C(O)R, —C(O)CH2C(O)R, —S(O)R, —S(O)2R,—C(O)N(Ra)(Rb), —SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or a C1-6 aliphaticgroup optionally substituted with R;

wherein each R, each Ra and each Rb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

m and n are each independently selected from the group consisting ofintegers 1.

279. A compound or a tautomer, a mesomer, a racemate, an enantiomer or adiastereoisomer thereof, or a mixture thereof, or a pharmaceuticallyacceptable salt thereof, wherein the compound comprises a structureshown as formula (II-Cx) or formula (II-Cy):

wherein, L is -La-Lb-Lc-;

-La- is selected from the group consisting of:

wherein W is —(C(Rwa)(Rwb))wn-, Y is —(OCH2CH2)yn-Oyp-, and Z is—(C(Rza)(Rzb))zn;

wherein wn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of W is independently replaced by-Cyr-, —N(Rwx)C(O)—, —C(O)N(Rwx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRwx-,—O—, —S—, —SO—, —SO2-, —P(Rwx)-, —P(═O)(Rwx)-, —N(Rwx)SO2-, —SO2N(Rwx)-,—C(═S)—, —C(═NRwx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

wherein yn is selected from the group consisting of integers 0, and ypis 0 or 1;

wherein zn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of Z is independently replaced by-Cyr-, —N(Rzx)C(O)—, —C(O)N(Rzx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRzx-,—O—, —S—, —SO—, —SO2-, —P(Rzx)-, —P(═O)(Rzx)-, —N(Rzx)SO2-, —SO2N(Rzx)-,—C(═S)—, —C(═NRzx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

-Cyr- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or independently substituted with no less than 1substituent Rcx;

wherein each Rwa, each Rwb, each Rza, each Rzb, each Rwx, each Rzx andeach Rcx are each independently hydrogen, protium, deuterium, tritium,halogen, —NO2, —CN, —ORr, —SRr, —N(Rra)(Rrb), —C(O)Rr, —CO2Rr,—C(O)C(O)Rr, —C(O)CH2C(O)Rr, —S(O)Rr, —S(O)2Rr, —C(O)N(Rra)(Rrb),—SO2N(Rra)(Rrb), —OC(O)Rr, —N(R)SO2Rr, or a C1-6 aliphatic groupoptionally substituted with Rr;

wherein each Rr, each Rra and each Rrb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

-Lb- represents a peptide residue consisting of 2 to 7 amino acids;

-Lc- is selected from the group consisting of:

wherein RL1 and RL2 are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO2,—CN, —OH, —SH, —NH2, —C(O)H, —CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H,—S(O)2H, —C(O)NH2, —SO2NH2, —OC(O)H, —N(H)SO2H and a C1-6 aliphaticgroup;

wherein, X1 is selected from the group consisting of: N, P, andsaturated or unsaturated C;

when X1 is saturated C, X1 is substituted with Rn;

when X1 is saturated C, ring A is selected from the group consisting of:3-10 membered saturated or partially unsaturated heterocyclyl, and 3-10membered saturated or partially unsaturated carbocyclyl, wherein ring Ais unsubstituted or substituted with no less than 1 substituent R1a;

or, when X1 is unsaturated C, ring A is selected from the groupconsisting of: 6-10 membered aryl, 5-8 membered heteroaryl, 3-10membered partially unsaturated heterocyclyl, and 3-10 membered partiallyunsaturated carbocyclyl, wherein ring A is unsubstituted or substitutedwith no less than 1 substituent R1b;

or, when X1 is N or P, ring A is selected from the group consisting of:5-8 membered heteroaryl and 3-10 membered saturated or partiallyunsaturated heterocyclyl, wherein ring A is unsubstituted or substitutedwith no less than 1 substituent R1c;

when ring A is selected from the group consisting of: 6-10 memberedaryl, 5-8 membered heteroaryl, and 3-10 membered saturated or partiallyunsaturated carbocyclyl, ring A is substituted with p L2, wherein L2 isnot Rn;

or, when ring A is 3-10 membered saturated or partially unsaturatedheterocyclyl, ring A is substituted with p L2, or ring A comprises qring-forming heteroatom X2, and X2 is used for direct or indirectlinking of a ligand;

X2 is selected from the group consisting of: N and P;

L2 is -R2-L3-, and R2 is used for direct or indirect linking of aligand;

L3 is —(C(R3a)(R3b))m-, wherein 0 or no less than 1 methylene unit of L3is independently replaced by —N(R4)C(O)—, —C(O)N(R4)-, —C(O)—, —OC(O)—,—C(O)O—, —NR4-, —O—, —S—, —SO—, —SO2-, —P(R4)-, —P(═O)(R4)-, —N(R4)SO2-,—SO2N(R4)-, —C(═S)—, —C(═NR4)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

R2 is selected from the group consisting of: —O—, —(R2a)N—, —S— and—P(═O)(R2a)-;

L1 is —(C(R5a)(R5b))n-, wherein 0 or no less than 1 methylene unit of L1is independently replaced by —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—,—C(O)O—, —NR6-, —O—, —S—, —SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-,—SO2N(R6)-, —C(═S)—, —C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

wherein each R1a, each R1b, each R1c, each R2a, each R3a, each R3b, eachR4, each R5a, each R5b, each R6 and each Rn are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO2, —CN, —OR, —SR,—N(R^(a))(Rb), —C(O)R, —CO2R, —C(O)C(O)R, —C(O)CH2C(O)R, —S(O)R,—S(O)2R, —C(O)N(Ra)(Rb), —SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or a C1-6aliphatic group optionally substituted with R;

wherein each R, each Ra and each Rb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

m and n are each independently selected from the group consisting ofintegers 0, and p and q are each independently selected from the groupconsisting of integers 1.

280. A compound or a tautomer, a mesomer, a racemate, an enantiomer or adiastereoisomer thereof, or a mixture thereof, or a pharmaceuticallyacceptable salt thereof, wherein the compound comprises a structureshown as formula (III-C):

wherein, L is -La-Lb-Lc-;

-La- is selected from the group consisting of:

wherein W is —(C(Rwa)(Rwb))wn-, Y is —(OCH2CH2)yn-Oyp, and Z is—(C(Rza)(Rzb))zn;

wherein wn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of W is independently replaced by-Cyr-, —N(Rwx)C(O)—, —C(O)N(Rwx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRwx-,—O—, —S—, —SO—, —SO2-, —P(Rwx)-, —P(═O)(Rwx)-, —N(Rwx)SO2-, —SO2N(Rwx)-,—C(═S)—, —C(═NRwx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

wherein yn is selected from the group consisting of integers 0, and ypis 0 or 1;

wherein zn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of Z is independently replaced by-Cyr-, —N(Rzx)C(O)—, —C(O)N(Rzx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRzx-,—O—, —S—, —SO—, —SO2-, —P(Rzx)-, —P(═O)(Rzx)-, —N(Rzx)SO2-, —SO2N(Rzx)-,—C(═S)—, —C(═NRzx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

-Cyr- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or independently substituted with no less than 1substituent Rcx;

wherein each Rwa, each Rwb, each Rza, each Rzb, each Rwx, each Rzx andeach Rcx are each independently hydrogen, protium, deuterium, tritium,halogen, —NO2, —CN, —ORr, —SRr, —N(Rra)(Rrb), —C(O)Rr, —CO2Rr,—C(O)C(O)Rr, —C(O)CH2C(O)Rr, —S(O)Rr, —S(O)2Rr, —C(O)N(Rra)(Rrb),—SO2N(Rra)(Rrb), —OC(O)Rr, —N(R)SO2Rr, or a C1-6 aliphatic groupoptionally substituted with Rr;

wherein each Rr, each Rra and each Rrb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

-Lb- represents a peptide residue consisting of 2 to 7 amino acids;

-Lc- is selected from the group consisting of:

wherein RL1 and RL2 are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO2,—CN, —OH, —SH, —NH2, —C(O)H, —CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H,—S(O)2H, —C(O)NH2, —SO2NH2, —OC(O)H, —N(H)SO2H and a C1-6 aliphaticgroup;

wherein R1 is selected from the group consisting of: —O—, —(R2)N—,—P(═O)(R2)- and —S—;

X is selected from the group consisting of: -L1-C(R1a)(R1b)—C(O)—,-L1-C(R1a)(R1b)—C(S)—, -L1-L0- and -L3-L2-;

L1 is —(C(R3a)(R3b))m-, wherein 0 or no less than 1 methylene unit of L1is independently replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or —C(═N2)-;

L0 is —C(R2a)(R2b)-, or L0 is —C(═S)—, —C(═NR4a)- or —C(═N2)-;

L2 is —C(R5a)(R5b)-, wherein 0 or 1 methylene unit of L2 is replaced by—N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—, —C(O)O—, —NR6-, —O—, —S—,—SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-, —SO2N(R6)-, —C(═S)—,—C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

L3 is —(C(R7a)(R7b))n-, wherein no less than 1 methylene unit of L3 isindependently replaced by —N(R8)C(O)—, —C(O)N(R8)-, —OC(O)—, —C(O)O—,—NRB—, —O—, —S—, —SO—, —SO2-, —P(R8)-, —P(═O)(R8)-, —N(R8)SO2-,—SO2N(R8)-, —N═N—, —C═N— or —N═C—, and 0 or no less than 1 methyleneunit of L3 is also independently replaced by —C(O)—, —C(═S)—, —C(═NR8)-or —C(═N2)-;

wherein each R1a, each Rib, each R2, each R2a, each R2b, each R3a, eachR3b, each R4a, each R4b, each R5a, each R5b, each R6, each R7a, each R7band each R8 are each independently hydrogen, protium, deuterium,tritium, halogen, —NO2, —CN, —OR, —SR, —N(Ra)(Rb), —C(O)R, —CO2R,—C(O)C(O)R, —C(O)CH2C(O)R, —S(O)R, —S(O)2R, —C(O)N(Ra)(Rb),—SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or a C1-6 aliphatic group optionallysubstituted with R;

wherein each R, each Ra and each Rb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

m is selected from the group consisting of integers 0, and n is selectedfrom the group consisting of integers 1;

when R1 is —O— or —HN— and X is -L1-CH2-C(O)—, no less than 1 methyleneunit of L1 is independently replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or—C(═N2)-, or each R3a and each R3b are not both hydrogen;

when R1 is —HN—, X is -L1-L0-, and L0 is —CH2-, no less than 1 methyleneunit of L1 is independently replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or—C(═N2)-, or each R3a and each R3b are not both hydrogen;

when R1 is —O—, X is -L3-C(O)—, and 1 methylene unit of L3 is replacedby —NRB, R8 is not —CH2-CH2-NH2;

when R1 is —NH—, and X is -L3-C(O)—, no less than 1 methylene unit of L3is replaced by —N(R8)C(O)—, —OC(O)—, —C(O)O—, —S—, —SO—, —SO2-, —P(R8)-,—P(═O)(R8)-, —N(R8)SO2-, —SO2N(R8)-, —N═N—, —C═N— or —N═C—.

281. A compound or a tautomer, a mesomer, a racemate, an enantiomer or adiastereoisomer thereof, or a mixture thereof, or a pharmaceuticallyacceptable salt thereof, wherein the compound comprises a structureshown as formula (I-D):

wherein, Ab is a ligand, and an average connection number Na is aninteger or a decimal from 1 to 10;

L is -La-Lb-Lc-;

-La- is selected from the group consisting of:

wherein W is —(C(Rwa)(Rwb))wn-, Y is —(OCH2CH2)yn-Oyp-, and Z is—(C(Rza)(Rzb))zn;

wherein wn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of W is independently replaced by-Cyr-, —N(Rwx)C(O)—, —C(O)N(Rwx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRwx-,—O—, —S—, —SO—, —SO2-, —P(Rwx)-, —P(═O)(Rwx)-, —N(Rwx)SO2-, —SO2N(Rwx)-,—C(═S)—, —C(═NRwx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

wherein yn is selected from the group consisting of integers 0, and ypis 0 or 1;

wherein zn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of Z is independently replaced by-Cyr-, —N(Rzx)C(O)—, —C(O)N(Rzx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRzx-,—O—, —S—, —SO—, —SO2-, —P(Rzx)-, —P(═O)(Rzx)-, —N(Rzx)SO2-, —SO2N(Rzx)-,—C(═S)—, —C(═NRzx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

-Cyr- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or independently substituted with no less than 1substituent Rcx;

wherein each Rwa, each Rwb, each Rza, each Rzb, each Rwx, each Rzx andeach Rcx are each independently hydrogen, protium, deuterium, tritium,halogen, —NO2, —CN, —ORr, —SRr, —N(Rra)(Rrb), —C(O)Rr, —CO2Rr,—C(O)C(O)Rr, —C(O)CH2C(O)Rr, —S(O)Rr, —S(O)2Rr, —C(O)N(Rra)(Rrb),—SO2N(Rra)(Rrb), —OC(O)Rr, —N(R)SO2Rr, or a C1-6 aliphatic groupoptionally substituted with Rr;

wherein each Rr, each Rra and each Rrb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

-Lb- represents a peptide residue consisting of 2 to 7 amino acids;

-Lc- is selected from the group consisting of:

wherein RL1 and RL2 are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO2,—CN, —OH, —SH, —NH2, —C(O)H, —CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H,—S(O)2H, —C(O)NH2, —SO2NH2, —OC(O)H, —N(H)SO2H and a C1-6 aliphaticgroup;

wherein R1 is selected from the group consisting of: —O—, —(R2)N—,—P(═O)(R2)- and —S—;

L2 is —(C(R3a)(R3b))m-R,

wherein 0 or no less than 1 methylene unit of L2 is independentlyreplaced by -Cy-, —N(R4)C(O)—, —C(O)N(R4)-, —C(O)—, —OC(O)—, —C(O)O—,—NR4-, —O—, —S—, —SO—, —SO2-, —P(R4)-, —P(═O)(R4)-, —N(R4)SO2-,—SO2N(R4)-, —C(═S)—, —C(═NR4)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

L1 is —(C(R5a)(R5b))n-,

wherein 0 or no less than 1 methylene unit of L1 is independentlyreplaced by -Cy-, —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—, —C(O)O—,—NR6-, —O—, —S—, —SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-,—SO2N(R6)-, —C(═S)—, —C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

-Cy- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein -Cy-is unsubstituted or independently substituted with no less than 1substituent R7;

wherein each R3a, each R3b, each R4, each R5a, each R5b and each R6 areeach independently hydrogen, protium, deuterium, tritium, halogen, —NO2,—CN, —OR, —SR, —N(Ra)(Rb), —C(O)R, —CO2R, —C(O)C(O)R, —C(O)CH2C(O)R,—S(O)R, —S(O)2R, —C(O)N(Ra)(Rb), —SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or aC1-6 aliphatic group optionally substituted with R; or, R3a and R5a, R4and R5a, R3a and R6 or R4 and R6 each independently optionally form aring B together with an atom therebetween, wherein the ring B isselected from the group consisting of: 5-8 membered heteroarylene and3-10 membered saturated or partially unsaturated heterocyclylene, andthe ring B is unsubstituted or substituted with no less than 1substituent R8;

wherein each R2, each R7 and each R8 are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OR, —SR, —N(Ra)(Rb),—C(O)R, —CO2R, —C(O)C(O)R, —C(O)CH2C(O)R, —S(O)R, —S(O)2R,—C(O)N(Ra)(Rb), —SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or a C1-6 aliphaticgroup optionally substituted with R;

wherein each R, each Ra and each Rb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

m and n are each independently selected from the group consisting ofintegers 1.

282. A compound or a tautomer, a mesomer, a racemate, an enantiomer or adiastereoisomer thereof, or a mixture thereof, or a pharmaceuticallyacceptable salt thereof, wherein the compound comprises a structureshown as formula (II-Dx) or formula (II-Dy):

wherein, Ab is a ligand, and an average connection number Na is aninteger or a decimal from 1 to 10;

L is -La-Lb-Lc-;

-La- is selected from the group consisting of:

wherein W is —(C(Rwa)(Rwb))wn-, Y is —(OCH2CH2)yn-Oyp-, and Z is—(C(Rza)(Rzb))zn;

wherein wn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of W is independently replaced by-Cyr-, —N(Rwx)C(O)—, —C(O)N(Rwx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRwx-,—O—, —S—, —SO—, —SO2-, —P(Rwx)-, —P(═O)(Rwx)-, —N(Rwx)SO2-, —SO2N(Rwx)-,—C(═S)—, —C(═NRwx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

wherein yn is selected from the group consisting of integers 0, and ypis 0 or 1;

wherein zn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of Z is independently replaced by-Cyr-, —N(Rzx)C(O)—, —C(O)N(Rzx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRzx-,—O—, —S—, —SO—, —SO2-, —P(Rzx)-, —P(═O)(Rzx)-, —N(Rzx)SO2-, —SO2N(Rzx)-,—C(═S)—, —C(═NRzx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

-Cyr- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or independently substituted with no less than 1substituent Rcx;

wherein each Rwa, each Rwb, each Rza, each Rzb, each Rwx, each Rzx andeach Rcx are each independently hydrogen, protium, deuterium, tritium,halogen, —NO2, —CN, —ORr, —SRr, —N(Rra)(Rrb), —C(O)Rr, —CO2Rr,—C(O)C(O)Rr, —C(O)CH2C(O)Rr, —S(O)Rr, —S(O)2Rr, —C(O)N(Rra)(Rrb),—SO2N(Rra)(Rrb), —OC(O)Rr, —N(R)SO2Rr, or a C1-6 aliphatic groupoptionally substituted with Rr;

wherein each Rr, each Rra and each Rrb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

-Lb- represents a peptide residue consisting of 2 to 7 amino acids;

-Lc- is selected from the group consisting of:

wherein RL1 and RL2 are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO2,—CN, —OH, —SH, —NH2, —C(O)H, —CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H,—S(O)2H, —C(O)NH2, —SO2NH2, —OC(O)H, —N(H)SO2H and a C1-6 aliphaticgroup;

wherein, X1 is selected from the group consisting of: N, P, andsaturated or unsaturated C;

when X1 is saturated C, X1 is substituted with Rn;

when X1 is saturated C, ring A is selected from the group consisting of:3-10 membered saturated or partially unsaturated heterocyclyl, and 3-10membered saturated or partially unsaturated carbocyclyl, wherein ring Ais unsubstituted or substituted with no less than 1 substituent R1a;

or, when X1 is unsaturated C, ring A is selected from the groupconsisting of: 6-10 membered aryl, 5-8 membered heteroaryl, 3-10membered partially unsaturated heterocyclyl, and 3-10 membered partiallyunsaturated carbocyclyl, wherein ring A is unsubstituted or substitutedwith no less than 1 substituent R1b;

or, when X1 is N or P, ring A is selected from the group consisting of:5-8 membered heteroaryl and 3-10 membered saturated or partiallyunsaturated heterocyclyl, wherein ring A is unsubstituted or substitutedwith no less than 1 substituent R1c;

when ring A is selected from the group consisting of: 6-10 memberedaryl, 5-8 membered heteroaryl, and 3-10 membered saturated or partiallyunsaturated carbocyclyl, ring A is substituted with p L2, wherein L2 isnot Rn;

or, when ring A is 3-10 membered saturated or partially unsaturatedheterocyclyl, ring A is substituted with p L2, or ring A comprises qring-forming heteroatom X2, and X2 is used for direct or indirectlinking of a ligand;

X2 is selected from the group consisting of: N and P;

L2 is -R2-L3-, and R2 is used for direct or indirect linking of aligand;

L3 is —(C(R3a)(R3b))m-, wherein 0 or no less than 1 methylene unit of L3is independently replaced by —N(R4)C(O)—, —C(O)N(R4)-, —C(O)—, —OC(O)—,—C(O)O—, —NR4-, —O—, —S—, —SO—, —SO2-, —P(R4)-, —P(═O)(R4)-, —N(R4)SO2-,—SO2N(R4)-, —C(═S)—, —C(═NR4)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

R2 is selected from the group consisting of: —O—, —(R2a)N—, —S— and—P(═O)(R2a)-;

L1 is —(C(R5a)(R5b))n-, wherein 0 or no less than 1 methylene unit of L1is independently replaced by —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—,—C(O)O—, —NR6-, —O—, —S—, —SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-,—SO2N(R6)-, —C(═S)—, —C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

wherein each R1a, each Rib, each R1c, each R2a, each R3a, each R3b, eachR4, each R5a, each R5b, each R6 and each Rn are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO2, —CN, —OR, —SR,—N(Ra)(Rb), —C(O)R, —CO2R, —C(O)C(O)R, —C(O)CH2C(O)R, —S(O)R, —S(O)2R,—C(O)N(Ra)(Rb), —SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or a C1-6 aliphaticgroup optionally substituted with R;

wherein each R, each Ra and each Rb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

m and n are each independently selected from the group consisting ofintegers 0, and p and q are each independently selected from the groupconsisting of integers 1.

283. A compound or a tautomer, a mesomer, a racemate, an enantiomer or adiastereoisomer thereof, or a mixture thereof, or a pharmaceuticallyacceptable salt thereof, wherein the compound comprises a structureshown as formula (III-D):

wherein, Ab is a ligand, and an average connection number Na is aninteger or a decimal from 1 to 10;

L is -La-Lb-Lc-;

-La- is selected from the group consisting of:

wherein W is —(C(Rwa)(Rwb))wn-, Y is —(OCH2CH2)yn-Oyp, and Z is—(C(Rza)(Rzb))zn;

wherein wn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of W is independently replaced by-Cyr-, —N(Rwx)C(O)—, —C(O)N(Rwx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRwx-,—O—, —S—, —SO—, —SO2-, —P(Rwx)-, —P(═O)(Rwx)-, —N(Rwx)SO2-, —SO2N(Rwx)-,—C(═S)—, —C(═NRwx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

wherein yn is selected from the group consisting of integers 0, and ypis 0 or 1;

wherein zn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of Z is independently replaced by-Cyr-, —N(Rzx)C(O)—, —C(O)N(Rzx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRzx-,—O—, —S—, —SO—, —SO2-, —P(Rzx)-, —P(═O)(Rzx)-, —N(Rzx)SO2-, —SO2N(Rzx)-,—C(═S)—, —C(═NRzx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

-Cyr- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or independently substituted with no less than 1substituent Rcx;

wherein each Rwa, each Rwb, each Rza, each Rzb, each Rwx, each Rzx andeach Rcx are each independently hydrogen, protium, deuterium, tritium,halogen, —NO2, —CN, —ORr, —SRr, —N(Rra)(Rrb), —C(O)Rr, —CO2Rr,—C(O)C(O)Rr, —C(O)CH2C(O)Rr, —S(O)Rr, —S(O)2Rr, —C(O)N(Rra)(Rrb),—SO2N(Rra)(Rrb), —OC(O)Rr, —N(R)SO2Rr, or a C1-6 aliphatic groupoptionally substituted with Rr;

wherein each Rr, each Rra and each Rrb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

-Lb- represents a peptide residue consisting of 2 to 7 amino acids;

-Lc- is selected from the group consisting of:

wherein RL1 and RL2 are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO2,—CN, —OH, —SH, —NH2, —C(O)H, —CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H,—S(O)2H, —C(O)NH2, —SO2NH2, —OC(O)H, —N(H)SO2H and a C1-6 aliphaticgroup;

wherein R1 is selected from the group consisting of: —O—, —(R2)N—,—P(═O)(R2)- and —S—;

X is selected from the group consisting of: -L1-C(R1a)(R1b)—C(O)—,-L1-C(R1a)(R1b)—C(S)—, -L1-L0- and -L3-L2-;

L1 is —(C(R3a)(R3b))m-, wherein 0 or no less than 1 methylene unit of L1is independently replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or —C(═N2)-;

L0 is —C(R2a)(R2b)-, or L0 is —C(═S)—, —C(═NR4a)- or —C(═N2)-;

L2 is —C(R5a)(R5b)-, wherein 0 or 1 methylene unit of L2 is replaced by—N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—, —C(O)O—, —NR6-, —O—, —S—,—SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-, —SO2N(R6)-, —C(═S)—,—C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

L3 is —(C(R7a)(R7b))n-, wherein no less than 1 methylene unit of L3 isindependently replaced by —N(R8)C(O)—, —C(O)N(R8)-, —OC(O)—, —C(O)O—,—NR6-, —O—, —S—, —SO—, —SO2-, —P(R8)-, —P(═O)(R8)—, —N(R8)SO2-,—SO2N(R8)—, —N═N—, —C═N— or —N═C—, and 0 or no less than 1 methyleneunit of L3 is also independently replaced by —C(O)—, —C(═S)—, —C(═NR8)-or —C(═N2)-;

wherein each R1a, each R1b, each R2, each R2a, each R2b, each R3a, eachR3b, each R4a, each R4b, each R5a, each R5b, each R6, each R7a, each R7band each R8 are each independently hydrogen, protium, deuterium,tritium, halogen, —NO2, —CN, —OR, —SR, —N(R^(a))(Rb), —C(O)R, —CO2R,—C(O)C(O)R, —C(O)CH2C(O)R, —S(O)R, —S(O)2R, —C(O)N(Ra)(Rb),—SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or a C1-6 aliphatic group optionallysubstituted with R;

wherein each R, each Ra and each Rb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

m is selected from the group consisting of integers 0, and n is selectedfrom the group consisting of integers 1;

when R1 is —O— or —HN— and X is -L1-CH2-C(O)—, no less than 1 methyleneunit of L1 is independently replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or—C(═N2)-, or each R3a and each R3b are not both hydrogen;

when R1 is —HN—, X is -L1-L0-, and L0 is —CH2-, no less than 1 methyleneunit of L1 is independently replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or—C(═N2)-, or each R3a and each R3b are not both hydrogen;

when R1 is —O—, X is -L3-C(O)—, and 1 methylene unit of L3 is replacedby —NRB, R8 is not —CH2-CH2-NH2;

when R1 is —NH—, and X is -L3-C(O)—, no less than 1 methylene unit of L3is replaced by —N(R8)C(O)—, —OC(O)—, —C(O)O—, —S—, —SO—, —SO2-, —P(R8)-,—P(═O)(R8)-, —N(R8)SO2-, —SO2N(R8)-, —N═N—, —C═N— or —N═C—.

284. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 281-283, wherein the ligand Ab is an antibody or anantigen-binding fragment thereof.

285. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof. or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 281-284, wherein the ligand Ab is selected from thegroup consisting of: a chimeric antibody, a humanized antibody and afully humanized antibody.

286. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 281-285, wherein the ligand Ab targets the following:HER2, HER3, B7H3, TROP2, Claudin 18.2, CD30, CD33, CD70 or EGFR.

287. A compound of general formula (I-E) or a tautomer, a mesomer, aracemate, an enantiomer or a diastereoisomer thereof, or a mixturethereof, or a pharmaceutically acceptable salt thereof,

wherein R1 is selected from the group consisting of: —O—, —(R2)N—,—P(═O)(R2)- and —S—;

L2 is —(C(R3a)(R3b))m-R,

wherein 0 or no less than 1 methylene unit of L2 is independentlyreplaced by -Cy-, —N(R4)C(O)—, —C(O)N(R4)-, —C(O)—, —OC(O)—, —C(O)O—,—NR4-, —O—, —S—, —SO—, —SO2-, —P(R4)-, —P(═O)(R4)-, —N(R4)SO2-,—SO2N(R4)-, —C(═S)—, —C(═NR4)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

L1 is —(C(R5a)(R5b))n-,

wherein 0 or no less than 1 methylene unit of L1 is independentlyreplaced by -Cy-, —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—, —C(O)O—,—NR6-, —O—, —S—, —SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-,—SO2N(R6)-, —C(═S)—, —C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

-Cy- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein -Cy-is unsubstituted or independently substituted with no less than 1substituent R7;

wherein each R3a, each R3b, each R4, each R5a, each R5b and each R6 areeach independently hydrogen, protium, deuterium, tritium, halogen, —NO2,—CN, —OR, —SR, —N(Ra)(Rb), —C(O)R, —CO2R, —C(O)C(O)R, —C(O)CH2C(O)R,—S(O)R, —S(O)2R, —C(O)N(Ra)(Rb), —SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or aC1-6 aliphatic group optionally substituted with R; or, R3a and R5a, R4and R5a, R3a and R6 or R4 and R6 each independently optionally form aring B together with an atom therebetween, wherein the ring B isselected from the group consisting of: 5-8 membered heteroarylene and3-10 membered saturated or partially unsaturated heterocyclylene, andthe ring B is unsubstituted or independently substituted with no lessthan 1 substituent R8;

wherein each R2, each R7 and each R8 are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OR, —SR, —N(Ra)(Rb),—C(O)R, —CO2R, —C(O)C(O)R, —C(O)CH2C(O)R, —S(O)R, —S(O)2R,—C(O)N(Ra)(Rb), —SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or a C1-6 aliphaticgroup optionally substituted with R;

wherein each R, each Ra and each Rb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

m and n are each independently selected from the group consisting ofintegers 1.

288. A compound of general formula (II-Ex) or (II-Ey), or a tautomer, amesomer, a racemate, an enantiomer or a diastereoisomer thereof, or amixture thereof, or a pharmaceutically acceptable salt thereof,

wherein, X1 is selected from the group consisting of: N, P, andsaturated or unsaturated C;

when X1 is saturated C, X1 is substituted with Rn;

when X1 is saturated C, ring A is selected from the group consisting of:3-10 membered saturated or partially unsaturated heterocyclyl, and 3-10membered saturated or partially unsaturated carbocyclyl, wherein ring Ais unsubstituted or independently substituted with no less than 1substituent R1a;

or, when X1 is unsaturated C, ring A is selected from the groupconsisting of: 6-10 membered aryl, 5-8 membered heteroaryl, 3-10membered partially unsaturated heterocyclyl, and 3-10 membered partiallyunsaturated carbocyclyl, wherein ring A is unsubstituted orindependently substituted with no less than 1 substituent Rib;

or, when X1 is N or P, ring A is selected from the group consisting of:5-8 membered heteroaryl and 3-10 membered saturated or partiallyunsaturated heterocyclyl, wherein ring A is unsubstituted orindependently substituted with no less than 1 substituent R1c;

when ring A is selected from the group consisting of: 6-10 memberedaryl, 5-8 membered heteroaryl, and 3-10 membered saturated or partiallyunsaturated carbocyclyl, ring A is substituted with p L2, wherein L2 isnot Rn;

or, when ring A is 3-10 membered saturated or partially unsaturatedheterocyclyl, ring A is substituted with p L2, or ring A comprises qring-forming heteroatom X2, and X2 is used for direct or indirectlinking of a ligand;

X2 is selected from the group consisting of: N and P;

L2 is -R2-L3-, and R2 is used for direct or indirect linking of aligand;

L3 is —(C(R3a)(R3b))m-, wherein 0 or no less than 1 methylene unit of L3is independently replaced by —N(R4)C(O)—, —C(O)N(R4)-, —C(O)—, —OC(O)—,—C(O)O—, —NR4-, —O—, —S—, —SO—, —SO2-, —P(R4)-, —P(═O)(R4)-, —N(R4)SO2-,—SO2N(R4)-, —C(═S)—, —C(═NR4)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

R2 is selected from the group consisting of: —O—, —(R2a)N—, —S— and—P(═O)(R2a)-;

L1 is —(C(R5a)(R5b))n-, wherein 0 or no less than 1 methylene unit of L1is independently replaced by —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—,—C(O)O—, —NR6-, —O—, —S—, —SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-,—SO2N(R6)-, —C(═S)—, —C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

wherein each R1a, each Rib, each R1c, each R2a, each R3a, each R3b, eachR4, each R5a, each R5b, each R6 and each Rn are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO2, —CN, —OR, —SR,—N(Ra)(Rb), —C(O)R, —CO2R, —C(O)C(O)R, —C(O)CH2C(O)R, —S(O)R, —S(O)2R,—C(O)N(Ra)(Rb), —SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or a C1-6 aliphaticgroup optionally substituted with R;

wherein each R, each Ra and each Rb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

m and n are each independently selected from the group consisting ofintegers 0, and p and q are each independently selected from the groupconsisting of integers 1.

289. A compound of general formula (III-E) or a tautomer, a mesomer, aracemate, an enantiomer or a diastereoisomer thereof, or a mixturethereof, or a pharmaceutically acceptable salt thereof,

wherein R1 is selected from the group consisting of: —O—, —(R2)N—,—P(═O)(R2)- and —S—;

X is selected from the group consisting of: -L1-C(R1a)(R1b)—C(O)—,-L1-C(R1a)(R1b)—C(S)—, -L1-L0- and -L3-L2-;

L1 is —(C(R3a)(R3b))m-, wherein 0 or no less than 1 methylene unit of L1is independently replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or —C(═N2)-;

L0 is —C(R2a)(R2b)-, or L0 is —C(═S)—, —C(═NR4a)- or —C(═N2)-;

L2 is —C(R5a)(R5b)-, wherein 0 or 1 methylene unit of L2 is replaced by—N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—, —C(O)O—, —NR6-, —O—, —S—,—SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-, —SO2N(R6)-, —C(═S)—,—C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

L3 is —(C(R7a)(R7b))n-, wherein no less than 1 methylene unit of L3 isindependently replaced by —N(R8)C(O)—, —C(O)N(R8)-, —OC(O)—, —C(O)O—,—NRB—, —O—, —S—, —SO—, —SO2-, —P(R8)-, —P(═O)(R8)-, —N(R8)SO2-,—SO2N(R8)-, —N═N—, —C═N— or —N═C—, and 0 or no less than 1 methyleneunit of L3 is also independently replaced by —C(O)—, —C(═S)—, —C(═NR8)-or —C(═N2)-;

wherein each R1a, each Rib, each R2, each R2a, each R2b, each R3a, eachR3b, each R4a, each R4b, each R5a, each R5b, each R6, each R7a, each R7band each R8 are each independently hydrogen, protium, deuterium,tritium, halogen, —NO2, —CN, —OR, —SR, —N(Ra)(Rb), —C(O)R, —CO2R,—C(O)C(O)R, —C(O)CH2C(O)R, —S(O)R, —S(O)2R, —C(O)N(Ra)(Rb),—SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or a C1-6 aliphatic group optionallysubstituted with R;

wherein each R, each Ra and each Rb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

m is selected from the group consisting of integers 0, and n is selectedfrom the group consisting of integers 1;

when R1 is —O— or —HN— and X is -L1-CH2-C(O)—, no less than 1 methyleneunit of L1 is independently replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or—C(═N2)-, or each R3a and each R3b are not both hydrogen;

when R1 is —HN—, X is -L1-L0-, and L0 is —CH2-, no less than 1 methyleneunit of L1 is independently replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or—C(═N2)-, or each R3a and each R3b are not both hydrogen;

when R1 is —O—, X is -L3-C(O)—, and 1 methylene unit of L3 is replacedby —NRB, R8 is not —CH2-CH2-NH2;

when R1 is —NH—, and X is -L3-C(O)—, no less than 1 methylene unit of L3is replaced by —N(R8)C(O)—, —OC(O)—, —C(O)O—, —S—, —SO—, —SO2-, —P(R8)-,—P(═O)(R8)-, —N(R8)SO2-, —SO2N(R8)-, —N═N—, —C═N— or —N═C—.

290. A compound of general formula (I-F) or a tautomer, a mesomer, aracemate, an enantiomer or a diastereoisomer thereof, or a mixturethereof, or a pharmaceutically acceptable salt thereof,

wherein, Lx is Lax-Lb-Lc-;

Lax-is selected from the group consisting of:

wherein Rhal is iodine or bromine;

wherein W is —(C(Rwa)(Rwb))wn-, Y is —(OCH2CH2)yn-Oyp-, and Z is—(C(Rza)(Rzb))zn;

wherein wn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of W is independently replaced by-Cyr-, —N(Rwx)C(O)—, —C(O)N(Rwx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRwx-,—O—, —S—, —SO—, —SO2-, —P(Rwx)-, —P(═O)(Rwx)-, —N(Rwx)SO2-, —SO2N(Rwx)-,—C(═S)—, —C(═NRwx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

wherein yn is selected from the group consisting of integers 0, and ypis 0 or 1;

wherein zn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of Z is independently replaced by-Cyr-, —N(Rzx)C(O)—, —C(O)N(Rzx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRzx-,—O—, —S—, —SO—, —SO2-, —P(Rzx)-, —P(═O)(Rzx)-, —N(Rzx)SO2-, —SO2N(Rzx)-,—C(═S)—, —C(═NRzx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

-Cyr- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or independently substituted with no less than 1substituent Rcx;

wherein each Rwa, each Rwb, each Rza, each Rzb, each Rwx, each Rzx andeach Rcx are each independently hydrogen, protium, deuterium, tritium,halogen, —NO2, —CN, —ORr, —SRr, —N(Rra)(Rrb), —C(O)Rr, —CO2Rr,—C(O)C(O)Rr, —C(O)CH2C(O)Rr, —S(O)Rr, —S(O)2Rr, —C(O)N(Rra)(Rrb),—SO2N(Rra)(Rrb), —OC(O)Rr, —N(R)SO2Rr, or a C1-6 aliphatic groupoptionally substituted with Rr;

wherein each Rr, each Rra and each Rrb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

-Lb- represents a peptide residue consisting of 2 to 7 amino acids;

-Lc- is selected from the group consisting of:

wherein RL1 and RL2 are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO2,—CN, —OH, —SH, —NH2, —C(O)H, —CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H,—S(O)2H, —C(O)NH2, —SO2NH2, —OC(O)H, —N(H)SO2H and a C1-6 aliphaticgroup;

wherein R1 is selected from the group consisting of: —O—, —(R2)N—,—P(═O)(R2)- and —S—;

L2 is —(C(R3a)(R3b))m-R,

wherein 0 or no less than 1 methylene unit of L2 is independentlyreplaced by -Cy-, —N(R4)C(O)—, —C(O)N(R4)-, —C(O)—, —OC(O)—, —C(O)O—,—NR4-, —O—, —S—, —SO—, —SO2-, —P(R4)-, —P(═O)(R4)-, —N(R4)SO2-,—SO2N(R4)-, —C(═S)—, —C(═NR4)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

L1 is —(C(R5a)(R5b))n-,

wherein 0 or no less than 1 methylene unit of L1 is independentlyreplaced by -Cy-, —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—, —C(O)O—,—NR6-, —O—, —S—, —SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-,—SO2N(R6)-, —C(═S)—, —C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

-Cy- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein -Cy-is unsubstituted or independently substituted with no less than 1substituent R7;

wherein each R3a, each R3b, each R4, each R5a, each R5b and each R6 areeach independently hydrogen, protium, deuterium, tritium, halogen, —NO2,—CN, —OR, —SR, —N(Ra)(Rb), —C(O)R, —CO2R, —C(O)C(O)R, —C(O)CH2C(O)R,—S(O)R, —S(O)2R, —C(O)N(Ra)(Rb), —SO2N(R^(a))(Rb), —OC(O)R, —N(R)SO2R,or a C1-6 aliphatic group optionally substituted with R; or, R3a andR5a, R4 and R5a, R3a and R6 or R4 and R6 each independently optionallyform a ring B together with an atom therebetween, wherein the ring B isselected from the group consisting of: 5-8 membered heteroarylene and3-10 membered saturated or partially unsaturated heterocyclylene, andthe ring B is unsubstituted or substituted with no less than 1substituent R8;

wherein each R2, each R7 and each R8 are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OR, —SR, —N(Ra)(Rb),—C(O)R, —CO2R, —C(O)C(O)R, —C(O)CH2C(O)R, —S(O)R, —S(O)2R,—C(O)N(Ra)(Rb), —SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or a C1-6 aliphaticgroup optionally substituted with R;

wherein each R, each Ra and each Rb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

m and n are each independently selected from the group consisting ofintegers 1.

291. A compound of general formula (II-Fx) or (II-Fy), or a tautomer, amesomer, a racemate, an enantiomer or a diastereoisomer thereof, or amixture thereof, or a pharmaceutically acceptable salt thereof,

wherein, Lx is Lax-Lb-Lc-;

Lax- is selected from the group consisting of:

wherein Rhal is iodine or bromine;

wherein W is —(C(Rwa)(Rwb))wn-, Y is —(OCH2CH2)yn-Oyp-, and Z is—(C(Rza)(Rzb))zn;

wherein wn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of W is independently replaced by-Cyr-, —N(Rwx)C(O)—, —C(O)N(Rwx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRwx-,—O—, —S—, —SO—, —SO2-, —P(Rwx)-, —P(═O)(Rwx)-, —N(Rwx)SO2-, —SO2N(Rwx)-,—C(═S)—, —C(═NRwx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

wherein yn is selected from the group consisting of integers 0, and ypis 0 or 1;

wherein zn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of Z is independently replaced by-Cyr-, —N(Rzx)C(O)—, —C(O)N(Rzx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRzx-,—O—, —S—, —SO—, —SO2-, —P(Rzx)-, —P(═O)(Rzx)-, —N(Rzx)SO2-, —SO2N(Rzx)-,—C(═S)—, —C(═NRzx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

-Cyr- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or independently substituted with no less than 1substituent Rcx;

wherein each Rwa, each Rwb, each Rza, each Rzb, each Rwx, each Rzx andeach Rcx are each independently hydrogen, protium, deuterium, tritium,halogen, —NO2, —CN, —ORr, —SRr, —N(Rra)(Rrb), —C(O)Rr, —CO2Rr,—C(O)C(O)Rr, —C(O)CH2C(O)Rr, —S(O)Rr, —S(O)2Rr, —C(O)N(Rra)(Rrb),—SO2N(Rra)(Rrb), —OC(O)Rr, —N(R)SO2Rr, or a C1-6 aliphatic groupoptionally substituted with Rr;

wherein each Rr, each Rra and each Rrb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

-Lb- represents a peptide residue consisting of 2 to 7 amino acids;

-Lc- is selected from the group consisting of:

wherein RL1 and RL2 are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO2,—CN, —OH, —SH, —NH2, —C(O)H, —CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H,—S(O)2H, —C(O)NH2, —SO2NH2, —OC(O)H, —N(H)SO2H and a C1-6 aliphaticgroup;

wherein, X1 is selected from the group consisting of: N, P, andsaturated or unsaturated C;

when X1 is saturated C, X1 is substituted with Rn;

when X1 is saturated C, ring A is selected from the group consisting of:3-10 membered saturated or partially unsaturated heterocyclyl, and 3-10membered saturated or partially unsaturated carbocyclyl, wherein ring Ais unsubstituted or substituted with no less than 1 substituent R1a;

or, when X1 is unsaturated C, ring A is selected from the groupconsisting of: 6-10 membered aryl, 5-8 membered heteroaryl, 3-10membered partially unsaturated heterocyclyl, and 3-10 membered partiallyunsaturated carbocyclyl, wherein ring A is unsubstituted or substitutedwith no less than 1 substituent R1b;

or, when X1 is N or P, ring A is selected from the group consisting of:5-8 membered heteroaryl and 3-10 membered saturated or partiallyunsaturated heterocyclyl, wherein ring A is unsubstituted or substitutedwith no less than 1 substituent R1c;

when ring A is selected from the group consisting of: 6-10 memberedaryl, 5-8 membered heteroaryl, and 3-10 membered saturated or partiallyunsaturated carbocyclyl, ring A is substituted with p L2, wherein L2 isnot Rn;

or, when ring A is 3-10 membered saturated or partially unsaturatedheterocyclyl, ring A is substituted with p L2, or ring A comprises qring-forming heteroatom X2, and X2 is used for direct or indirectlinking of a ligand;

X2 is selected from the group consisting of: N and P;

L2 is -R2-L3-, and R2 is used for direct or indirect linking of aligand;

L3 is —(C(R3a)(R3b))m-, wherein 0 or no less than 1 methylene unit of L3is independently replaced by —N(R4)C(O)—, —C(O)N(R4)-, —C(O)—, —OC(O)—,—C(O)O—, —NR4-, —O—, —S—, —SO—, —SO2-, —P(R4)-, —P(═O)(R4)-, —N(R4)SO2-,—SO2N(R4)-, —C(═S)—, —C(═NR4)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

R2 is selected from the group consisting of: —O—, —(R2a)N—, —S— and—P(═O)(R2a)-;

L1 is —(C(R5a)(R5b))n-, wherein 0 or no less than 1 methylene unit of L1is independently replaced by —N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—,—C(O)O—, —NR6-, —O—, —S—, —SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-,—SO2N(R6)-, —C(═S)—, —C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

wherein each R1a, each Rib, each R1c, each R2a, each R3a, each R3b, eachR4, each R5a, each R5b, each R6 and each Rn are each independentlyhydrogen, protium, deuterium, tritium, halogen, —NO2, —CN, —OR, —SR,—N(Ra)(Rb), —C(O)R, —CO2R, —C(O)C(O)R, —C(O)CH2C(O)R, —S(O)R, —S(O)2R,—C(O)N(Ra)(Rb), —SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or a C1-6 aliphaticgroup optionally substituted with R;

wherein each R, each Ra and each Rb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

m and n are each independently selected from the group consisting ofintegers 0, and p and q are each independently selected from the groupconsisting of integers 1.

292. A compound of general formula (III-F) or a tautomer, a mesomer, aracemate, an enantiomer or a diastereoisomer thereof, or a mixturethereof, or a pharmaceutically acceptable salt thereof,

wherein, Lx is Lax-Lb-Lc-;

Lax- is selected from the group consisting of:

wherein Rhal is iodine or bromine;

wherein W is —(C(Rwa)(Rwb))wn-, Y is —(OCH2CH2)yn-Oyp, and Z is—(C(Rza)(Rzb))zn;

wherein wn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of W is independently replaced by-Cyr-, —N(Rwx)C(O)—, —C(O)N(Rwx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRwx-,—O—, —S—, —SO—, —SO2-, —P(Rwx)-, —P(═O)(Rwx)-, —N(Rwx)SO2-, —SO2N(Rwx)-,—C(═S)—, —C(═NRwx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

wherein yn is selected from the group consisting of integers 0, and ypis 0 or 1;

wherein zn is selected from the group consisting of integers 0, and

0 or no less than 1 methylene unit of Z is independently replaced by-Cyr-, —N(Rzx)C(O)—, —C(O)N(Rzx)-, —C(O)—, —OC(O)—, —C(O)O—, —NRzx-,—O—, —S—, —SO—, —SO2-, —P(Rzx)-, —P(═O)(Rzx)-, —N(Rzx)SO2-, —SO2N(Rzx)-,—C(═S)—, —C(═NRzx)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

-Cyr- is selected from the group consisting of: 6-10 membered arylene,5-8 membered heteroarylene, 3-10 membered heterocyclylene, and 3-10membered saturated or partially unsaturated carbocyclylene, wherein-Cyr- is unsubstituted or independently substituted with no less than 1substituent Rcx;

wherein each Rwa, each Rwb, each Rza, each Rzb, each Rwx, each Rzx andeach Rcx are each independently hydrogen, protium, deuterium, tritium,halogen, —NO2, —CN, —ORr, —SRr, —N(Rra)(Rrb), —C(O)Rr, —CO2Rr,—C(O)C(O)Rr, —C(O)CH2C(O)Rr, —S(O)Rr, —S(O)2Rr, —C(O)N(Rra)(Rrb),—SO2N(Rra)(Rrb), —OC(O)Rr, —N(R)SO2Rr, or a C1-6 aliphatic groupoptionally substituted with Rr;

wherein each Rr, each Rra and each Rrb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

-Lb- represents a peptide residue consisting of 2 to 7 amino acids;

-Lc- is selected from the group consisting of:

wherein RL1 and RL2 are each independently selected from the groupconsisting of: hydrogen, protium, deuterium, tritium, halogen, —NO2,—CN, —OH, —SH, —NH2, —C(O)H, —CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H,—S(O)2H, —C(O)NH2, —SO2NH2, —OC(O)H, —N(H)SO2H and a C1-6 aliphaticgroup;

wherein R1 is selected from the group consisting of: —O—, —(R2)N—,—P(═O)(R2)- and —S—;

X is selected from the group consisting of: -L1-C(R1a)(R1b)—C(O)—,-L1-C(R1a)(R1b)—C(S)—, -L1-L0- and -L3-L2-;

L1 is —(C(R3a)(R3b))m-, wherein 0 or no less than 1 methylene unit of L1is independently replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or —C(═N2)-;

L0 is —C(R2a)(R2b)-, or L0 is —C(═S)—, —C(═NR4a)- or —C(═N2)-;

L2 is —C(R5a)(R5b)-, wherein 0 or 1 methylene unit of L2 is replaced by—N(R6)C(O)—, —C(O)N(R6)-, —C(O)—, —OC(O)—, —C(O)O—, —NR6-, —O—, —S—,—SO—, —SO2-, —P(R6)-, —P(═O)(R6)-, —N(R6)SO2-, —SO2N(R6)-, —C(═S)—,—C(═NR6)-, —N═N—, —C═N—, —N═C— or —C(═N2)-;

L3 is —(C(R7a)(R7b))n-, wherein no less than 1 methylene unit of L3 isindependently replaced by —N(R8)C(O)—, —C(O)N(R8)-, —OC(O)—, —C(O)O—,—NRB—, —O—, —S—, —SO—, —SO2-, —P(R8)-, —P(═O)(R8)-, —N(R8)SO2-,—SO2N(R8)-, —N═N—, —C═N— or —N═C—, and 0 or no less than 1 methyleneunit of L3 is also independently replaced by —C(O)—, —C(═S)—, —C(═NR8)-or —C(═N2)-;

wherein each R1a, each Rib, each R2, each R2a, each R2b, each R3a, eachR3b, each R4a, each R4b, each R5a, each R5b, each R6, each R7a, each R7band each R8 are each independently hydrogen, protium, deuterium,tritium, halogen, —NO2, —CN, —OR, —SR, —N(Ra)(Rb), —C(O)R, —CO2R,—C(O)C(O)R, —C(O)CH2C(O)R, —S(O)R, —S(O)2R, —C(O)N(Ra)(Rb),—SO2N(Ra)(Rb), —OC(O)R, —N(R)SO2R, or a C1-6 aliphatic group optionallysubstituted with R;

wherein each R, each Ra and each Rb are each independently hydrogen,protium, deuterium, tritium, halogen, —NO2, —CN, —OH, —SH, —NH2, —C(O)H,—CO2H, —C(O)C(O)H, —C(O)CH2C(O)H, —S(O)H, —S(O)2H, —C(O)NH2, —SO2NH2,—OC(O)H, —N(H)SO2H or a C1-6 aliphatic group;

m is selected from the group consisting of integers 0, and n is selectedfrom the group consisting of integers 1;

when R1 is —O— or —HN— and X is -L1-CH2-C(O)—, no less than 1 methyleneunit of L1 is independently replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or—C(═N2)-, or each R3a and each R3b are not both hydrogen;

when R1 is —HN—, X is -L1-L0-, and L0 is —CH2-, no less than 1 methyleneunit of L1 is independently replaced by —C(O)—, —C(═S)—, —C(═NR4b)- or—C(═N2)-, or each R3a and each R3b are not both hydrogen;

when R1 is —O—, X is -L3-C(O)—, and 1 methylene unit of L3 is replacedby —NRB, R8 is not —CH2-CH2-NH2;

when R1 is —NH—, and X is -L3-C(O)—, no less than 1 methylene unit of L3is replaced by —N(R8)C(O)—, —OC(O)—, —C(O)O—, —S—, —SO—, —SO2-, —P(R8)-,—P(═O)(R8)-, —N(R8)SO2-, —SO2N(R8)-, —N═N—, —C═N— or —N═C—.

293. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 290-292, wherein Lax- is

294. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 290-293, wherein Lax-Lb-Lc- is selected from the groupconsisting of:

295. The compound of formula (I-B) according to any one of technicalschemes 266 and 269-277, the compound of formula (I-C) according totechnical scheme 278, the compound of formula (I-D) according to any oneof technical schemes 281 and 284-286, the compound of formula (I-E)according to technical scheme 287 or the compound of formula (I-F)according to any one of technical schemes 290 and 293-294, or thetautomer, the mesomer, the racemate, the enantiomer or thediastereoisomer thereof, or the mixture thereof, or the pharmaceuticallyacceptable salt thereof, wherein in a structure

comprised therein, L1, L2 and R1 are L1, L2 and R1, respectively, of thecompound according to any one of technical schemes 1-93.

296. The compound of formula (II-Bx) or (II-By) according to any one oftechnical schemes 267 and 269-277, the compound of formula (II-Cx) or(II-Cy) according to technical scheme 279, the compound of formula(II-Dx) or (II-Dy) according to any one of technical schemes 282 and284-286, the compound of formula (II-Ex) or (II-Ey) according totechnical scheme 288, or the compound of formula (II-Fx) or (II-Fy)according to any one of technical schemes 291 and 293-294, or thetautomer, the mesomer, the racemate, the enantiomer or thediastereoisomer thereof, or the mixture thereof, or the pharmaceuticallyacceptable salt thereof, wherein in a structure

comprised therein, L1, X1, A and L2 are L1, X1, A and L2, respectively,of the compound according to any one of technical schemes 94-167, or ina structure

comprised therein, L1, X1, A and X2 are L1, X1, A and X2, respectively,of the compound according to any one of technical schemes 94-167.

297. The compound of formula (III-B) according to any one of technicalschemes 268-277, the compound of formula (III-C) according to technicalscheme 280, the compound of formula (III-D) according to any one oftechnical schemes 283-286, the compound of formula (III-E) according totechnical scheme 289, or the compound of formula (III-F) according toany one of technical schemes 292-294, or the tautomer, the mesomer, theracemate, the enantiomer or the diastereoisomer thereof, or the mixturethereof, or the pharmaceutically acceptable salt thereof, wherein in astructure

comprised therein, R1 and X1 are R1 and X1, respectively, of thecompound according to any one of technical schemes 168-265.

298. The compound of formula (I-C) according to any one of technicalschemes 278 and 295 or the compound of formula (I-D) according to anyone of technical schemes 281, 284-286 and 295, or the tautomer, themesomer, the racemate, the enantiomer or the diastereoisomer thereof, orthe mixture thereof, or the pharmaceutically acceptable salt thereof,wherein in a structure -L- comprised therein, L is L of the compoundaccording to any one of technical schemes 266 and 269-277.

299. The compound of formula (II-Cx) or (II-Cy) according to any one oftechnical schemes 279 and 296 or the compound of formula (II-Dx) or(II-Dy) according to any one of technical schemes 282, 284-286 and 296,or the tautomer, the mesomer, the racemate, the enantiomer or thediastereoisomer thereof, or the mixture thereof, or the pharmaceuticallyacceptable salt thereof, wherein in a structure -L- comprised therein, Lis L of the compound according to any one of technical schemes 267 and269-277.

300. The compound of formula (III-C) according to any one of technicalschemes 280 and 297 or the compound of formula (III-D) according to anyone of technical schemes 283-286 and 297, or the tautomer, the mesomer,the racemate, the enantiomer or the diastereoisomer thereof, or themixture thereof, or the pharmaceutically acceptable salt thereof,wherein in a structure -L- comprised therein, L is L of the compoundaccording to any one of technical schemes 268-277.

301. The compound of formula (I-F) according to any one of technicalschemes 290, 293-294 and 295, or the tautomer, the mesomer, theracemate, the enantiomer or the diastereoisomer thereof, or the mixturethereof, or the pharmaceutically acceptable salt thereof, wherein Lb andLc are Lb and Lc, respectively, of the compound according to any one oftechnical schemes 266 and 269-277.

302. The compound of formula (II-Fx) or (II-Fy) according to any one oftechnical schemes 291, 293-294 and 296, or the tautomer, the mesomer,the racemate, the enantiomer or the diastereoisomer thereof, or themixture thereof, or the pharmaceutically acceptable salt thereof,wherein Lb and Lc are Lb and Lc, respectively, of the compound accordingto any one of technical schemes 267 and 269-277.

303. The compound of formula (III-F) according to any one of technicalschemes 292-294 and 297, or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof, wherein Lb and Lc are Lband Lc, respectively, of the compound according to any one of technicalschemes 268-277.

304. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-303, comprising the ligand-drug conjugate or apharmaceutically acceptable salt or a solvate thereof.

305. A method for preparing the compound or the tautomer, the mesomer,the racemate, the enantiomer or the diastereoisomer thereof, or themixture thereof, or the pharmaceutically acceptable salt thereofaccording to any one of technical schemes 1-304, comprising linking acytotoxic drug to a ligand Ab via the structure of formula (I-B)according to any one of technical schemes 266, 269-277 and 295.

306. A method for preparing the compound or the tautomer, the mesomer,the racemate, the enantiomer or the diastereoisomer thereof, or themixture thereof, or the pharmaceutically acceptable salt thereofaccording to any one of technical schemes 1-304, comprising linking acytotoxic drug to a ligand Ab via the structure of formula (II-Bx) or(II-By) according to any one of technical schemes 267, 269-277 and 296.

307. A method for preparing the compound or the tautomer, the mesomer,the racemate, the enantiomer or the diastereoisomer thereof, or themixture thereof, or the pharmaceutically acceptable salt thereofaccording to any one of technical schemes 1-304, comprising linking acytotoxic drug to a ligand Ab via the structure of formula (III-B)according to any one of technical schemes 268, 269-277 and 297.

308. A pharmaceutical composition, comprising the compound or thetautomer, the mesomer, the racemate, the enantiomer or thediastereoisomer thereof, or the mixture thereof, or the pharmaceuticallyacceptable salt thereof according to any one of technical schemes 1-304,and a pharmaceutically acceptable carrier.

309. Use of the compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to any one oftechnical schemes 1-304, and/or the pharmaceutical composition accordingto technical scheme 308, in preparing a medicament for treating and/orpreventing a tumor.

310. The use according to technical scheme 309, wherein the tumor isselected from the group consisting of tumors associated with expressionof the following: HER2, HER3, B7H3, TROP2, Claudin 18.2, CD30, CD33,CD70 and EGFR.

311. The use according to any one of technical schemes 309-310, whereinthe tumor is selected from the group consisting of: lung cancer, kidneycancer, urinary tract carcinoma, colorectal cancer, prostatic cancer,glioblastoma multiforme, ovarian cancer, pancreatic cancer, breastcancer, melanoma, liver cancer, bladder cancer, stomach cancer andesophageal cancer.

Without being bound by any theory, the following examples are intendedonly to illustrate the compounds, preparation methods, use, etc., of thepresent application, and are not intended to limit the scope of thepresent application.

EXAMPLES

The structure of the compounds is determined by nuclear magneticresonance (NMR) or mass spectrometry (MS). NMR is performed using a Quantum-I NMR spectrometer with deuterated dimethyl sulfoxide (DMSO-D),deuterated chloroform (CDCl3), and deuterated methanol (CD3OD) assolvents, and tetramethylsilane (TMS) as an internal standard, andchemical shifts are given in units of 10_6(ppm).

MS is performed using an Angilent 6230 ESI-TOF mass spectrometer(manufacturer: Agilent, type c: 6230)

UPLC is performed using a Waters AcquityUPLCSQD liquidchromatograph-mass spectrometer (Poroshell 120 EC-C18, 2.1 mm×50 mm, 1.9μm column).

HPLC is performed using an Agilent 1260 high-performance liquidchromatograph (TOSOH G3000 SW SEC column).

UV is measured using a Thermo Nanodrop 2000 spectrophotometer.

The proliferation inhibition rate and the IC5Q value are measured usingan EnVision microplate reader (PerkinElmer).

Yantai Yellow Sea HSGF254 or Qingdao GF254 silica gel plate is adoptedas a thin layer chromatography (TLC) silica gel plate. The specificationadopted by the TLC is 0.15-0.20 mm, and the specification adopted by thethin layer chromatography for the separation and purification ofproducts is 0.4-0.5 mm.

Yantai Yellow Sea silica gel of 200-300 mesh is generally utilized as acarrier in column chromatography.

Known starting materials of the present disclosure can be synthesizedusing or according to methods known in the art, or can be purchased fromcompanies such as ABCR GMBH & Co. KG, Acros Organnics, Aldrich ChemicalCompany, Accela ChemBio Inc., and Darui Chemicals.

In the examples, all reactions are carried out under an argon atmosphereor a nitrogen atmosphere unless otherwise stated.

The argon atmosphere or nitrogen atmosphere is created by connecting thereaction flask to an argon or nitrogen balloon with a volume of about 1L.

The hydrogen atmosphere is created by connecting a reaction flask to ahydrogen balloon with a volume of about 1 L.

In the examples, the solution in the reaction is an aqueous solutionunless otherwise stated.

In the examples, the reaction temperature is room temperature unlessotherwise stated. The room temperature is the optimum reactiontemperature, which ranges from 20° C. to 30° C.

The system of eluents for column chromatography and the system ofdeveloping agents for thin layer chromatography used for purifyingcompounds include: A: dichloromethane and isopropanol system, B:dichloromethane and methanol system, and C: petroleumether and ethylacetate system. The volume ratio of solvents is regulated according todifferent polarities of the compound, and can also be regulated byadding a small amount of triethylamine and acidic or alkaline reagent.

Some of the compounds of the present disclosure are characterized byTOF-LC/MS. TOF-LC/MS is performed using Agilent 6230 time-of-flight massspectrometer and Agilent 1290-Infinity ultra high performance liquidchromatograph.

Example 1 Preparation of Compounds Preparation Example 1.1.(1s,4R)—N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-4-hydroxycyclohexane-1-carb oxamide

Step 1.

DIEA (486 mg, 3.76 mmol) was added to a solution of KI4 (1.00 g, 1.88mmol), HATU (691 mg, 1.88 mmol) and 1 a (350 mg, 1.88 mmol) in DMF (18mL) at 0° C. under nitrogen atmosphere, and the mixture was stirred at25° C. for 3 h. After the starting material was consumed completely asdetected by TLC (EA), the reaction solution was added dropwise todeionized water (320 mL) and filtered to give a gray solid (1.02 g,yield 90%).

Step 2.

NaHCO₃ (417 mg, 5.00 mmol) as a solid was added to a solution of 1b (100mg, 0.166 mmol) in MeOH/DCM (1/1, 5 mL), and the mixture was stirred at25° C. for 3 h. After the reaction was completed as detected by TLC(EA), the reaction solution was filtered, dried by rotary evaporation atlow temperature, slurried with aq. HCl (0.5 M, 10 mL), filtered andpurified by prep-HPLC (0.1% TFA) to give a yellow solid (14 mg).

MS m/z (ESI): 562 [M+1]

H-NMR (400 MHz, DMSO-D): 8.37 (d, 1H), 7.81 (d, 1H), 7.32 (s, 1H),5.57-5.55 (m, 1H), 5.43 (s, 2H), 5.18 (dd, 2H), 3.79 (m, 1H), 3.19-3.17(m, 2H), 2.41 (s, 3H), 2.30-2.12 (m, 3H), 1.95-1.80 (m, 4H), 1.73-1.62(m, 2H), 1.59-1.32 (m, 4H), 0.89 (t, 3H)

Preparation Example 1.2.(1r,4S)—N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-4-hydroxycyclohexane-1-carboxamide

Step 1.

DIEA (486 mg, 3.76 mmol) was added to a solution of KI4 (1.00 g, 1.88mmol), HATU (691 mg, 1.88 mmol) and 2a (390 mg, 2.07 mmol) in DMF (18mL) at 0° C. under nitrogen atmosphere, and the mixture was stirred at25° C. for 3 h. After the starting material was consumed completely asdetected by TLC (EA), the reaction solution was added dropwise todeionized water (320 mL) and filtered to give a gray solid (913 mg,yield 81%).

Step 2.

NaHCO₃ (42 mg, 0.50 mmol) as a solid was added to a solution of 2b (100mg, 0.166 mmol) in MeOH/DCM (1/1, 3 mL), and the mixture was stirred at25° C. for 3 h. After the reaction was completed as detected by TLC(EA), the reaction solution was filtered, dried by rotary evaporation atlow temperature, slurried with aq. HCl (0.5 M, 10 mL), filtered andpurified by prep-HPLC (0.1% TFA) to give a gray solid (18 mg, yield20%).

MS m/z (ESI): 562 [M+1]

H-NMR (400 MHz, DMSO-D): 8.39 (d, 1H), 7.78 (d, 1H), 7.29 (s, 1H), 6.52(s, 1H), 5.53-5.51 (m, 1H), 5.41 (s, 2H), 5.12 (dd, 2H), 4.57 (d, 1H),3.17-3.14 (m, 2H), 2.38 (s, 3H), 2.13-2.07 (m, 3H), 1.90-1.70 (m, 6H),1.52-1.41 (m, 2H), 1.18-1.01 (m, 2H), 0.86 (t, 3H)

Preparation Example 1.3.(1s,3R)—N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxycyclobutane-1-carboxamide

Step 1:

DIEA (500 mg, 3.87 mmol) was added to a solution of KI4 (900 mg, 1.69mmol), HATU (691 mg, 1.88 mmol) and 3a (320 mg, 2.00 mmol) in DMF (18mL) at 0° C. under nitrogen atmosphere, and the mixture was stirred at25° C. for 3 h. After the starting material was consumed completely asdetected by TLC (EA), the reaction solution was added dropwise todeionized water (320 mL) and filtered to give a gray solid (850 mg,yield 87%).

Step 2.

NaHCO₃ (42 mg, 0.50 mmol) as a solid was added to a solution of 3b (100mg, 0.174 mmol) in MeOH/DCM (1/1, 3 mL), and the mixture was stirred at25° C. for 3 h. After the reaction was completed as detected by TLC(EA), the reaction solution was filtered, dried by rotary evaporation atlow temperature, slurried with aq. HCl (0.5 M, 10 mL), filtered andpurified by prep-HPLC (0.1% TFA), and then lyophilized to give a graysolid (15 mg, yield 16%).

MS m/z (ESI): 534 [M+1]

H-NMR (400 MHz, DMSO-D): 8.45 (d, 1H), 7.81 (d, 1H), 7.32 (s, 1H), 6.52(m, 1H), 5.58-5.56 (m, 1H), 5.44 (s, 2H), 5.14 (dd, 2H), 3.96 (m, 1H),3.48 (m, 1H), 3.19 (m, 2H), 2.53-2.28 (m, 3H), 2.48 (s, 3H), 2.20-2.00(m, 4H), 1.95-1.80 (m, 2H), 0.89 (t, 3H)

Preparation Example 1.4. (1r,3S)—N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxycyclobutane-1-carboxamide

Step 1.

DIEA (486 mg, 3.76 mmol) was added to a solution of KI4 (1.00 g, 1.88mmol), HATU (691 mg, 1.88 mmol) and 4a (310 mg, 1.88 mmol) in DMF (18mL) at 0° C. under nitrogen atmosphere, and the mixture was stirred at25° C. for 3 h. After the starting material was consumed completely asdetected by TLC (EA), the reaction solution was added dropwise todeionized water (320 mL) and filtered to give a gray solid (910 mg,yield 84%).

Step 2.

NaHCO₃ (42 mg, 0.50 mmol) as a solid was added to a solution of 4b (100mg, 1.58 mmol) in MeOH/DCM (1/1, 3 mL), and the mixture was stirred at25° C. for 3 h. After the reaction was completed as detected by TLC(EA), the reaction solution was filtered, dried by rotary evaporation atlow temperature, slurried with aq. HCl (0.5 M, 10 mL), filtered andpurified by prep-HPLC (0.1% TFA), and then lyophilized to give a yellowsolid (13 mg, yield 14%).

MS m/z (ESI): 534 [M+1]

H-NMR (400 MHz, DMSO-D): 8.39 (d, 1H), 7.77 (d, 1H), 7.29 (s, 1H), 6.52(m, 1H), 5.58-5.54 (m, 1H), 5.41 (s, 2H), 5.18-5.06 (m, 3H), 4.39-4.33(m, 1H), 3.19-3.07 (m, 2H), 2.97-2.82 (m, 1H), 2.49-2.36 (m, 2H), 2.38(s, 3H), 2.20-1.96 (m, 4H), 1.93-1.79 (m, 2H), 0.87 (t, 3H)

Preparation Example 1.5(1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((R)-3-hydroxy-2-oxopyrrolidin-1-yl)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3′,4′:6,7]indolizino[[1,2-b]quinoline-10,13-dione](1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((S)-3-hydroxy-2-oxopyrrolidin-1-yl)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3′,4′:6,7]indolizino[[1,2-b]quinoline-10,13-dione]

Step 1.

TsOH (300 mg, 7.46 mmol) was added slowly to a solution of 5a (20.0 g,149 mmol) in 2,2-dimethoxypropane (500 mL) at 25° C., and the mixturewas stirred for 17 h. After the reaction was completed as detected byTLC (EA), the reaction solution was directly purified by columnchromatography (PE:EA=1:0 to 1:1) to give a white solid (20 g, yield77%).

Step 2.

Ethanethiol (6.40 g, 103 mmol), DCC (30.9 g, 149 mmol) and DMAP (280 mg,2.30 mmol) were added to a solution of 5b (20.0 g, 115 mmol) in DCM (500mL) at 0° C. After the addition was completed, the reaction solution waswarmed to 25° C. and stirred for 17 h. After the reaction was completedas detected by TLC (PE/EA=2/1), the reaction solution was filtered andwashed with saturated brine (100 mL). The organic phase was dried,filtered, concentrated and purified by column chromatography (PE:EA=1:0to 10:1) to give a pale yellow oil (20 g, yield 80%).

Step 3.

Pd(OAc)₂ (1.00 g, 4.58 mmol) and Et₃SiH (5.33 g, 45.8 mmol) were addedto a solution of 5c (5.00 g, 22.9 mmol) in acetone (100 mL) at 0° C.,and the mixture was stirred at 25° C. for 3 h. After the reaction wascompleted as detected by TLC (PE/EA=2/1), the reaction solution wasfiltered, concentrated and purified by column chromatography (PE:EA=1:0to 1:1) to give a yellow oil (3.6 g, yield 82%).

Step 4.

HOAc (6.8 mg, 0.11 mmol) was added to a solution of 5d (15 mg, 71 μmol),a solution of KI4 (40 mg, 71 μmol) in DCE (4 mL), and DMA (1 mL), andthe mixture was stirred at 25° C. for 20 min. Sodium borohydride acetate(24 mg, 0.11 mmol) was added to the above reaction solution, and themixture was stirred at 25° C. for 4 h. After the reaction was completedas detected by LCMS, the reaction solution was added with saturatedbrine, extracted with DCM/MeOH (5:1, 20 mL) three times, washed withsaturated aqueous sodium chloride solution (30 mL), dried over anhydroussodium sulfate, dried by rotary evaporation, and purified by prep-HPLC(0.1% TFA) to give two products: P10A (8 mg, RT: 0.895 min, yield 20%)as a yellow solid and P10B (9 mg, RT: 0.917 min, yield 23%) as a yellowsolid.

P-I-1 AMS m/z (ESI): 520 [M+1]

P-I-1 A H-NMR (400 MHz, DMSO-D): 7.83 (d, 1H), 7.33 (s, 1H), 6.55 (m,1H), 5.60-5.58 (m, 1H), 5.44 (s, 2H), 5.01 (dd, 2H), 4.29 (t, 1H),3.31-3.11 (m, 4H), 2.96-2.82 (m, 1H), 2.42 (s, 3H), 2.29-2.12 (m, 2H),1.99-1.81 (m, 3H), 0.89 (t, 3H)

P-I-1B MS m/z (ESI): 520 [M+1]

P-I-1B H-NMR (400 MHz, DMSO-D): 7.84 (d, 1H), 7.33 (s, 1H), 6.55 (m,1H), 5.67-5.65 (m, 1H), 5.44 (s, 2H), 5.17 (dd, 2H), 4.28 (t, 1H),3.30-3.11 (m, 4H), 2.87 (t, 1H), 2.32-2.12 (m, 2H), 2.41 (s, 3H),1.95-1.82 (m, 2H), 1.78-1.65 (m, 1H), 0.90 (t, 3H)

Preparation Example 1.6.N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-mercaptopropanamide

DIEA (122 mg, 0.943 mmol) was added to a solution of KI4 (200 mg, 0.377mmol), HATU (214.7 mg, 0.565 mmol) and 6a (60 mg, 0.565 mmol) in DMF (6mL) under nitrogen atmosphere, and the mixture was stirred at 25° C. for3 h. After the starting material was consumed completely as detected byLCMS, the reaction solution was added dropwise to water (90 mL) andfiltered to give a yellow solid (326 mg), which was prepared byprep-HPLC (0.5% TFA) to give a yellow solid (19 mg, yield 9.6%).

MS m/z (ESI): 524 [M+1]

H-NMR (400 MHz, DMSO-D): 8.55 (d, 1H), 7.83 (d, 1H), 7.33 (s, 1H), 6.54(m, 1H), 5.65-5.55 (m, 1H), 5.45 (s, 2H), 5.26 (dd, 2H), 3.23-3.17 (m,2H), 2.80-2.71 (dd, 2H), 2.70-2.66 (m, 2H), 2.43 (s, 3H), 2.19-2.14 (m,1H), 1.92-1.83 (m, 2H), 0.89 (t, 3H)

Preparation Example 1.7.(R)—N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3‘,4’: 6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide

DIEA (61 mg, 0.47 mmol) was added to a solution of KI4 (100 mg, 0.188mmol), HATU (86 mg, 0.23 mmol) and 7a (22 mg, 0.21 mmol) in DMF (2 mL)under nitrogen atmosphere. After the addition was completed, the mixturewas reacted at 25° C. for 2.5 h. After the starting material wasconsumed completely as detected by LCMS, the reaction solution was addedto water (20 mL), and a solid was precipitated. The resulting mixturewas filtered to give the product (13 mg, yield 11%).

MS m/z (ESI): 522 [M+1]

H-NMR (400 MHz, DMSO-D): 8.44 (d, 1H), 7.80 (d, 1H), 7.32 (s, 1H), 6.54(m, 1H), 5.60-5.50 (m, 1H), 5.44 (s, 2H), 5.22 (dd, 2H), 4.10-4.00 (m,1H), 3.30-3.17 (m, 2H), 2.41 (s, 3H), 2.38-2.10 (m, 4H), 1.96-1.80 (m,2H), 1.11 (d, 3H), 0.89 (t, 3H)

Preparation Example 1.8.N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxy-3-methylbutanamide

A solution of DIEA (61 mg, 0.47 mmol) in DMF (1 mL) was added to asolution of KI4 (100 mg, 0.188 mmol), HATU (86 mg, 0.23 mmol) and 8a (24mg, 0.21 mmol) in DMF (1 mL) under nitrogen atmosphere. After theaddition was completed, the mixture was reacted at 25° C. for 3 h. Afterthe starting material was consumed completely as detected by TLC (EA),the reaction solution was purified by preparative chromatography to givea yellow solid (17 mg, yield 17%).

MS m/z (ESI): 536 [M+1]

H-NMR (400 MHz, DMSO-D): 8.42 (d, 1H), 7.79 (d, 1H), 7.30 (s, 1H), 6.53(m, 1H), 5.59-5.55 (m, 1H), 5.42 (s, 2H), 5.22 (dd, 2H), 4.69 (s, 1H),3.20-3.11 (m, 2H), 2.39 (sc, 3H), 2.29 (s, 2H), 2.20-2.08 (m, 2H), 1.19(d, 6H), 0.87 (t, 3H)

Preparation Example 1.9.N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-1-hydroxycyclobutane-1-carboxamide(reference example 1)

Step 1.

KI4 (50 mg, 0.094 mmol), 25a (11 mg, 0.094 mmol) and HATU (39 mg, 0.103mmol) were added to DMF (3 mL), and DIEA (30 mg, 0.235 mmol) was addedafter purging with nitrogen. The mixture was reacted at 25° C. for 1.5h. After the reaction was completed as detected by LCMS, the reactionsolution was added dropwise to water (50 mL) with stirring. After theaddition was completed, the reaction solution was left to stand for 5min and filtered, and the filter cake was lyophilized to give 25 (20 mg,yield 40%) as a gray solid.

MS-ESI: m/z 534.3 [M+H]+.

1H NMR (400 MHz, DMSO-d6) δ 8.26 (d, J=9.1 Hz, 1H), 7.72 (d, J=10.9 Hz,1H), 7.28 (s, 1H), 6.51 (s, 1H), 6.12 (s, 1H), 5.59-5.50 (m, 1H), 5.40(s, 2H), 5.15 (d, J=18.8 Hz, 1H), 4.98 (d, J=19.0 Hz, 1H), 3.28-3.16 (m,1H), 3.15-3.02 (m, 1H), 2.74-2.52 (m, 2H), 2.36 (s, 3H), 2.24-2.04 (m,4H), 1.92-1.79 (m, 4H), 0.86 (t, J=7.3 Hz, 3H).

Preparation Example 1.10. N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7] indolino[1,2-b]quinolin-1-yl)-3-aminopropionamide (referenceexample 2)

Step 1.

KI4 (50 mg, 0.094 mmol), 26a (18 mg, 0.094 mmol) and HATU (39 mg, 0.103mmol) were added to DMF (3 mL), and DIEA (48 mg, 0.376 mmol) was addedafter purging with nitrogen. The mixture was reacted at 25° C. for 1.5h. After the reaction was completed as detected by LCMS, the reactionsolution was added dropwise to water (50 mL) with stirring. After theaddition was completed, the reaction solution was left to stand for 5min and filtered, and the filter cake was lyophilized to give a graysolid 26b, (30 mg, yield 52%). MS-ESI: m/z 607.4 [M+H]+.

Step 2.

26b (30 mg, 0.049 mmol) was dissolved in DCM (2 mL), and the solutionwas purged with N2, cooled to 0° C. and added with TFA (0.5 mL). Themixture was reacted at 0° C. for 1.5 h. After the reaction was completedas detected by LCMS, the reaction solution was dried by rotaryevaporation at low temperature, washed with DCM once, added withacetonitrile and water and lyophilized to give a yellow solid (20 mg,yield 80%).

MS-ESI: m/z 507.1 [M+H]+.

1H NMR (400 MHz, DMSO-d6) δ 8.67 (d, J=8.6 Hz, 1H), 7.86-7.66 (m, 4H),7.32 (s, 1H), 6.56 (brs, 1H), 5.63-5.54 (m, 1H), 5.43 (s, 2H), 5.29 (d,J=18.9 Hz, 1H), 5.22 (d, J=18.9 Hz, 1H), 3.23-3.15 (m, 2H), 3.13-3.03(m, 2H), 2.57-2.51 (m, 2H), 2.43-2.38 (m, 3H), 2.27-2.08 (m, 2H),1.94-1.78 (m, 2H), 0.87 (t, J=7.3 Hz, 3H).

Preparation example 1.11.N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxypropanamide(reference example 3)

Step 1.

KI4 (100 mg, 0.188 mmol), 11a (33 mg, 0.188 mmol) and HATU (77 mg, 0.203mmol) were added to DMF (3 mL), and DIEA (73 mg, 0.564 mmol) was addedafter purging with nitrogen. The mixture was reacted at 25° C. for 1.5h. After the reaction was completed as detected by LCMS, the reactionsolution was added dropwise to water (50 mL) with stirring. After theaddition was completed, the reaction solution was left to stand for 5min and filtered, and the filter cake was lyophilized to give a graysolid 11b, (80 mg, yield 72%). MS-ESI: m/z 592.4 [M+H]+.

Step 2.

27b (80 mg, 0.135 mmol) was dissolved in DCM (2 mL), and the solutionwas purged with N2, cooled to 0° C. and added with TFA (0.5 mL). Themixture was reacted at 0° C. for 1.5 h. After the reaction was completedas detected by LCMS, the reaction solution was dried by rotaryevaporation at low temperature, purified by PTLC, added withacetonitrile and water and lyophilized to give a white solid (25 mg,yield 36%).

MS-ESI: m/z 508.3 [M+H]+.

1H NMR (400 MHz, DMSO-d6) δ 8.44 (d, J=8.6 Hz, 1H), 7.79 (d, J=11.0 Hz,1H), 7.30 (s, 1H), 6.53 (s, 1H), 5.60-5.53 (m, 1H), 5.42 (s, 2H),5.28-5.15 (m, 2H), 4.59 (t, J=5.1 Hz, 1H), 3.72-3.63 (m, 2H), 3.21-3.12(m, 2H), 2.40 (s, 3H), 2.32 (t, J=6.4 Hz, 2H), 2.21-2.06 (m, 2H),1.92-1.79 (m, 2H), 0.87 (t, J=7.3 Hz, 3H).

Preparation Example 1.12.N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxy-2-methylbutanamide(reference example 4)

Step 1.

12a (13 mg, 0.113 mmol), KI4 (60 mg, 0.113 mmol) and HATU (50 mg, 0.135mmol) were added to a 25 mL three-necked flask and dissolved with DMF (2mL), followed by the slow addition of DIEA (44 mg, 0.339 mmol). Afterthe reaction was completed as detected by LCMS, the reaction solutionwas purified by reverse-phase column chromatography to give a yellowpowder (37.2 mg, yield 62%).

MS-ESI: m/z 536.3 [M+H]+.

1H NMR (400 MHz, DMSO-d6) δ 8.50-8.23 (m, 1H), 7.82-7.75 (m, 1H),7.32-7.28 (m, 1H), 6.66-6.38 (m, 1H), 5.61-5.50 (m, 1H), 5.42 (s, 2H),5.31-5.10 (m, 2H), 3.77-3.61 (m, 2H), 3.22-3.12 (m, 2H), 2.56-2.52 (m,1H), 2.42-2.37 (m, 3H), 2.31-2.02 (m, 3H), 1.93-1.78 (m, 2H), 1.28-0.96(m, 7H), 0.91-0.84 (m, 3H).

Preparation Example 1.13.N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxycyclohexane-1-carboxamide

Step 1.

13a (22 mg, 0.15 mmol), KI4 (90 mg, 0.17 mmol) and HATU (68 mg, 0.18mmol) were added to a 25 mL three-necked flask and dissolved with DMF (2mL), followed by the slow addition of DIEA (58 mg, 0.45 mmol). After thereaction was completed as detected by LCMS, the reaction solution waspurified by reverse-phase column chromatography to give the compoundP-II-22 (52 mg, yield 55%).

MS-ESI: m/z 562.3 [M+H]+.

1H NMR (400 MHz, DMSO-d6) δ 8.45-8.32 (m, 1H), 7.78 (dd, J=11.0, 3.5 Hz,1H), 7.29 (d, J=2.2 Hz, 1H), 6.52 (s, 1H), 5.59-5.49 (m, 1H), 5.42 (s,2H), 5.27-5.01 (m, 2H), 3.24-3.08 (m, 2H), 2.71-2.54 (m, 1H), 2.39 (s,3H), 2.29-2.04 (m, 3H), 1.95-1.61 (m, 6H), 1.48-0.98 (m, 4H), 0.87 (t,J=7.3 Hz, 3H).

Preparation Example 1.14.N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-4-hydroxybenzamide-1-carboxamide

Step 1.

14a (31 mg, 0.169 mmol), KI4 (100 mg, 0.188 mmol) and HATU (78 mg, 0.206mmol) were added to a 25 mL three-necked flask and dissolved with DMF (4mL), followed by the slow addition of DIEA (67 mg, 0.516 mmol). Afterthe reaction was completed as detected by LCMS, the reaction solutionwas purified by reverse-phase column chromatography to give the compound14b (40 mg, yield 35%). MS-ESI: m/z 600.2 [M+H]+.

Step 2.

14b (40 mg, 0.067 mmol) was added to a 25 mL three-necked flask, andfound to be insoluble after the addition of ethyl acetate (30 mL) purgedwith hydrochloric acid gas, and then dissolved after the dropwiseaddition of methanol (2 mL). The solution was stirred for 1 h, and driedby rotary evaporation in vacuum under a water pump. After the reactionwas completed as detected by LCMS, the reaction solution was purified byreverse-phase column chromatography to give the compound P-II-23 (9 mg,yield 24%).

MS-ESI: m/z 556.3 [M+H]+.

1H NMR (400 MHz, DMSO-d6) δ 10.03 (s, 1H), 8.76 (d, J=8.5 Hz, 1H),7.85-7.77 (m, 3H), 7.30 (s, 1H), 6.80 (d, J=8.8 Hz, 2H), 6.52 (s, 1H),5.78-5.72 (m, 1H), 5.37 (s, 2H), 5.19 (d, J=18.8 Hz, 1H), 5.09 (d,J=19.0 Hz, 1H), 3.27-3.10 (m, 2H), 2.43-2.38 (m, 3H), 2.30-2.18 (m, 2H),1.90-1.77 (m, 2H), 0.85 (t, J=7.3 Hz, 3H).

Preparation Example 1.15.N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-5-hydroxypicolinamide

Step 1.

MOMBr (195 mg, 1.6 mmol) was added to an ice bath-cooled solution of 15a(200 mg, 1.3 mmol) and DIEA (336 mg, 2.6 mmol) in THF (6.5 mL) under N₂atmosphere. The reaction solution was stirred at 0° C. for 3 h. Afterthe reaction was completed, the reaction solution was added with MeOH(10 mL) at 0° C. and stirred for 20 min. The reaction solution wasconcentrated to give a pale yellow oil, which was purified by silica gelcolumn chromatography (PE/EA=20/1-10/1) to give 15b (240 mg, yield 93%)as a pale yellow solid.

Step 2.

NaOH (1 M in water, 2.2 mL, 2.2 mmol) was added dropwise to a solutionof 15b (220 mg, 1.1 mmol) in THF (5.0 mL) in an ice-water bath under N₂atmosphere, and the reaction solution was stirred at the sametemperature for 1.5 h. After the reaction was completed, the reactionsolution was adjusted to pH 2-3 at 0° C. with diluted hydrochloric acid(1 M in water), and concentrated to give a yellow oil, which wasdissolved with DCM (10 mL) and filtered and the filter cake was washedwith DCM (15 mL). The filtrate was concentrated to give 15c (205 mg) asa pale yellow oil, which was directly used in the next step.

Step 3.

HATU (173 mg, 0.46 mmol) was added to a suspension of 15c (76 mg, 0.42mmol), KI4 (185 mg, 0.35 mmol) and DIEA (136 mg, 1.05 mmol) in DMF (4.0mL) at 0° C. under N₂ atmosphere. The reaction solution was stirred for3 h to give a clear reaction solution. After the reaction was completedas detected by TLC, the reaction was added dropwise into water (10 mL),and a large amount of yellow solid was precipitated. The resultingmixture was filtered to give 15d (85 mg, yield 40%) as a yellow solid.

Step 4.

31d (80 mg, 0.13 mmol) was dissolved in a mixed solution of TFA/DCM(2/1) in an ice-water bath under N₂ atmosphere, and the reactionsolution was stirred for 0.5 h, concentrated, separated by prep-HPLC,and lyophilized to give P-II-24 (22 mg, yield 31%) as a yellow powder.

MS-ESI: m/z 557.3 [M+H]+.

1H NMR (400 MHz, DMSO-d6) δ 10.75 (brs, 1H), 9.24 (d, J=9.1 Hz, 1H),8.14 (d, J=2.7 Hz, 1H), 8.02 (d, J=8.6 Hz, 1H), 7.77 (d, J=10.9 Hz, 1H),7.37 (dd, J=8.6, 2.7 Hz, 1H), 7.28 (s, 1H), 5.76-5.66 (m, 1H), 5.34 (s,2H), 5.18 (d, J=19.1 Hz, 1H), 5.04 (d, J=19.0 Hz, 1H), 3.32-3.24 (m,1H), 3.19-3.08 (m, 1H), 2.38 (s, 3H), 2.36-2.20 (m, 2H), 1.92-1.75 (m,2H), 0.85 (t, J=7.3 Hz, 3H).

Preparation Example 1.16.N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-methylaminopropanamide

Step 1.

KI4 (100 mg, 0.188 mmol), 16a (42 mg, 0.207 mmol) and HATU (86 mg, 0.226mmol) were added to DMF (2 mL), followed by the addition of DIEA (73 mg,0.564 mmol), and the mixture was reacted at room temperature for 1 h.After the reaction was completed as detected by LCMS, the reaction wasadded dropwise to water (20 mL), and a solid was precipitated. Theresulting mixture was filtered, and the filter cake was washed withwater (20 mL×2) and lyophilized to give 32b (100 mg, yield 86%) as agray powder. MS-ESI: m/z 621.3 [M+H]+.

Step 2.

16b (25 mg, 0.040 mmol) was added to DCM (2 mL), and the mixture wascooled to 0° C., added with TFA (0.5 mL) and reacted at 0° C. for 1.5 h.After the reaction was completed as detected by LCMS, the reactionsolution was dried by rotary evaporation at low temperature to removeDCM and TFA, and the residue was added with water (10 mL) and MeCN (2mL) and lyophilized to give P-III-9 (23 mg, yield 90%) as a yellowpowder.

MS-ESI: m/z 521.3 [M+H]+.

1H NMR (400 MHz, DMSO-d6) δ 8.71 (d, J=8.6 Hz, 1H), 8.43 (brs, 2H), 7.81(d, J=11.0 Hz, 1H), 7.31 (s, 1H), 6.55 (brs, 1H), 5.58 (dt, J=8.8, 4.5Hz, 1H), 5.42 (s, 2H), 5.28 (d, J=18.9 Hz, 1H), 5.21 (d, J=18.9 Hz, 1H),3.25-3.12 (m, 4H), 2.62-2.53 (m, 5H), 2.40 (s, 3H), 2.26-2.06 (m, 2H),1.94-1.78 (m, 2H), 0.87 (t, J=7.3 Hz, 3H).

Preparation Example 1.17.N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-2-mercaptoacetamide

Step 1.

17a (118 mg, 0.353 mmol), KI4 (200 mg, 0.376 mmol) and HATU (160 mg,0.421 mmol) were added to a 25 mL three-necked flask and dissolved withDMF (6 mL), followed by the slow addition of DIEA (133 mg, 1.03 mmol).After the reaction was completed as detected by LCMS, the reactionsolution was slowly poured into water (50 mL), and the resulting mixturewas filtered to give a white solid, which was dried in vacuum to givethe product 17b (200 mg, yield 71%). MS-ESI: m/z 752.4 [M+H]+.

Step 2.

17b (200 mg, 0.266 mmol) was added to a 25 mL three-necked flask, andfound to be insoluble after the addition of ethyl acetate (50 mL) purgedwith hydrochloric acid gas, and then dissolved after the dropwiseaddition of methanol (5 mL). The solution was stirred for 1 h, and driedby rotary evaporation in vacuum under a water pump. After the reactionwas completed as detected by LCMS, the reaction solution was purified byreverse-phase column chromatography to give the compound P-III-1 (14 mg,yield 10%).

MS-ESI: m/z 510.1 [M+H]+.

1H NMR (400 MHz, DMSO-d6) δ 8.67-8.59 (m, 1H), 7.81 (d, J=11.0 Hz, 1H),7.33-7.29 (m, 1H), 6.53 (brs, 1H), 5.54 (dt, J=8.7, 4.5 Hz, 1H), 5.43(s, 2H), 5.30 (d, J=19.0 Hz, 1H), 5.20 (d, J=19.0 Hz, 1H), 3.20-3.09 (m,4H), 2.86 (t, J=8.1 Hz, 1H), 2.41 (s, 3H), 2.25-2.05 (m, 2H), 1.93-1.77(m, 2H), 0.91-0.83 (m, 3H).

Preparation Example 1.18.N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxyhexanamide

Step 1.

KI4 (50 mg, 0.094 mmol), 18a (14 mg, 0.106 mmol) and HATU (43 mg, 0.113mmol) were added to DMF (1 mL), followed by the addition of DIEA (36 mg,0.278 mmol), and the mixture was reacted at room temperature for 1 h.After the reaction was completed as detected by LCMS, the reactionsolution was added dropwise to water (10 mL), and a solid wasprecipitated. The resulting mixture was filtered, purified bypreparative chromatography and lyophilized to give P-III-27 (18 mg,yield 35%) as a yellow solid.

MS-ESI: m/z 550.3 [M+H]+.

1H NMR (400 MHz, DMSO-d6) δ 8.40 (dd, J=8.7, 2.5 Hz, 1H), 7.79 (d,J=10.7 Hz, 1H), 7.30 (d, J=1.2 Hz, 1H), 6.53 (brs, 1H), 5.60-5.52 (m,1H), 5.42 (s, 2H), 5.30-5.15 (m, 2H), 3.92-3.83 (m, 1H), 3.23-3.10 (m,2H), 2.44-2.36 (m, 3H), 2.23 (t, J=6.6 Hz, 2H), 2.20-2.03 (m, 2H),1.94-1.78 (m, 2H), 1.41-1.19 (m, 4H), 0.92-0.77 (m, 6H).

Preparation Example 1.19.N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-4-hydroxy-3-oxobutanamide

Step 1.

KI4 (50 mg, 0.094 mmol) and 19a (28.2 mg, 0.282 mmol) were dissolved inDMA (4 mL), followed by the addition of DIEA (60.7 mg, 0.470 mmol). Themixture was reacted at 20° C. for 1 h under nitrogen atmosphere, andthen warmed to 80° C. and reacted for 17 h. After the reaction wascompleted as detected by LCMS, the product MS-18 was obtained, which wasthen purified by preparative chromatography to give P-III-22 (10 mg,yield 20%) as a yellow solid.

MS-ESI: m/z 518.2 [M+H-H₂O]+.

1H NMR (400 MHz, DMSO-d6) δ 7.97 (s, 1H), 7.82 (d, J=10.9 Hz, 1H), 7.32(s, 1H), 6.54 (brs, 1H), 5.42 (s, 2H), 5.36 (d, J=19.0 Hz, 1H),5.24-5.11 (m, 2H), 5.03 (s, 1H), 4.66 (s, 2H), 3.31-3.06 (m, 3H),2.43-2.37 (m, 3H), 2.35-2.23 (m, 1H), 2.22-2.08 (m, 1H), 1.93-1.79 (m,2H), 0.87 (t, J=7.3 Hz, 3H).

Preparation Example 1.20.(S)-3-amino-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3‘,4’: 6,7]indolizino[1,2-b]quinolin-1-yl)butanamide

Step 1.

KI4 (100 mg, 0.188 mmol) and 20a (42 mg, 0.207 mmol) were dissolved inDMF (5 mL), followed by the addition of DIEA (73 mg, 0.564 mmol). Themixture was cooled to 0° C. under nitrogen atmosphere, added with HATU(93 mg, 0.244 mmol) and reacted at 0° C. for 2 h. After the startingmaterial was consumed completely as detected by TLC (EA/MeOH=10/1), thereaction solution was added to water (60 mL), and a solid wasprecipitated. The resulting mixture was filtered, and the solid waslyophilized to give 20b (89 mg, yield 76%) as a yellow solid. MS-ESI:m/z 621.3 [M+H]+.

Step 2.

20b (40 mg, 0.0644 mmol) was dissolved in DCM (2 mL), and the mixturewas cooled to 0° C. under nitrogen atmosphere, added dropwise with TFA(0.5 mL) and reacted at 0° C. for 1.5 h after the addition wascompleted. After the starting material was consumed completely asdetected by LCMS, the reaction solution was dried by rotary evaporation,added with acetonitrile and water and lyophilized to give P-III-28 (28.8mg, yield 86%) as a yellow solid.

MS-ESI: m/z 521.3 [M+H]+.

1H NMR (400 MHz, DMSO-d6) δ 8.74 (d, J=8.5 Hz, 1H), 7.90-7.77 (m, 4H),7.32 (s, 1H), 6.54 (brs, 1H), 5.57 (dt, J=9.0, 4.6 Hz, 1H), 5.42 (s,2H), 5.28 (d, J=18.9 Hz, 1H), 5.19 (d, J=18.9 Hz, 1H), 3.62-3.53 (m,2H), 3.24-3.12 (m, 2H), 2.48-2.38 (m, 4H), 2.25-2.07 (m, 2H), 1.94-1.79(m, 2H), 1.28-1.18 (m, 4H), 0.87 (t, J=7.3 Hz, 3H).

Preparation Example 1.21.(2S,4R)—N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-4-hydroxypyrrolidine-2-carboxamide

Step 1.

KI4 (100 mg, 0.188 mmol) and 21a (48 mg, 0.207 mmol) were dissolved inDMF (5 mL), followed by the addition of DIEA (73 mg, 0.564 mmol). Themixture was cooled to 0° C. under nitrogen atmosphere, added with HATU(93 mg, 0.244 mmol) and reacted at 0° C. for 2 h. After the reaction wascompleted as detected by TLC (EA/MeOH=10/1), the reaction was added towater (60 mL), and a solid was precipitated. The resulting mixture wasfiltered, and the solid was lyophilized to give 21b (72 mg, yield 59%)as a yellow solid. MS-ESI: m/z 649.3 [M+H]+.

Step 2.

21b (60 mg, 0.0925 mmol) was dissolved in DCM (2 mL), and the mixturewas cooled to 0° C. under nitrogen atmosphere, added dropwise with TFA(0.5 mL) and reacted at 0° C. for 1.5 h after the addition wascompleted. After the starting material was consumed completely asdetected by LCMS, the reaction solution was dried by rotary evaporation,added with acetonitrile and water and lyophilized to give P-II-25 (45.8mg, yield 90%) as a yellow solid.

MS-ESI: m/z 5493 [M+H]+.

1H NMR (400 MHz, DMSO-d6) δ 9.65 (brs, 1H), 9.14 (d, J=8.6 Hz, 1H), 8.85(brs, 1H), 7.82 (d, J=10.9 Hz, 1H), 7.32 (s, 1H), 6.54 (brs, 1H), 5.62(dt, J=9.0, 4.7 Hz, 1H), 5.42 (s, 2H), 5.27 (d, J=18.7 Hz, 1H), 5.09 (d,J=18.7 Hz, 1H), 4.48-4.41 (m, 1H), 4.39-4.27 (m, 1H), 3.26-3.08 (m, 3H),2.41 (s, 3H), 2.31-2.13 (m, 3H), 2.02-1.92 (m, 1H), 1.92-1.79 (m, 2H),1.30-1.20 (m, 2H), 0.88 (t, J=7.3 Hz, 3H).

Preparation Example 1.22.(R)-3-amino-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-4-hydroxybutanamide

Step 1.

22a (200 mg, 1.68 mmol) was dissolved in saturated NaHCO₃ solution (4mL) and THF (1 mL). The solution was reacted overnight. After theproduct was generated as detected by LCMS, the product was purified byreverse-phase column chromatography and lyophilized to give 22b (200 mg,59%) as a white solid. MS-ESI: m/z 203.6 [M+H]+.

Step 2.

22b (76 mg, 0.375 mmol), KI4 (200 mg, 0.375 mmol) and HATU (170 mg,0.450 mmol) were added to a 25 mL three-necked flask and dissolved withDMF (4 mL), followed by the slow addition of DIEA (145 mg, 1.125 mmol).After the reaction was completed as detected by LCMS, the reactionsolution was purified by reverse-phase column chromatography andlyophilized to give the compound 22c (40 mg, yield 17%). MS-ESI: m/z621.2 [M+H]+.

Step 3.

Compound 22c (40 mg, 0.0645 mmol) and Pd (PPh₃)₄ (22 mg, 0.0194 mmol)were added to a 25 mL three-necked flask and dissolved with THF (4 mL).The solution was added dropwise with N-methylmorpholine (0.2 mL) undernitrogen atmosphere, and reacted for 0.5 h. After the reaction wascompleted as detected by LCMS, the reaction solution was purified byreverse-phase column chromatography (ACN in water from 30%-70%), andlyophilized to give P-III-29 (14.3 mg, yield 41%) as a yellow powder.

MS-ESI: m/z 537.2 [M+H]+.

1H NMR (400 MHz, DMSO-d6) δ 8.68 (d, J=8.5 Hz, 1H), 7.95-7.82 (m, 3H),7.81 (d, J=10.9 Hz, 1H), 7.32 (s, 1H), 6.55 (s, 1H), 5.56 (dt, J=8.8,4.5 Hz, 1H), 5.42 (s, 2H), 5.30 (d, J=18.9 Hz, 1H), 5.22 (d, J=18.9 Hz,1H), 3.60 (dd, J=10.6, 3.6 Hz, 1H), 3.55-3.45 (m, 3H), 3.22-3.13 (m,2H), 2.56-2.52 (m, 1H), 2.48-2.44 (m, 1H), 2.44-2.37 (m, 3H), 2.26-2.05(m, 2H), 1.95-1.77 (m, 2H), 0.87 (t, J=7.3 Hz, 3H).

Preparation Example 1.23.(S)—N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide

Step 1.

DIEA (60.6 mg, 0.47 mmol) was added dropwise to a solution of KI4 (100mg, 0.19 mmol), HATU (85.7 mg, 0.23 mmol) and 23a (21.5 mg, 0.21 mmol)in DMF (2 mL) under nitrogen atmosphere. After the addition wascompleted, the mixture was stirred at 0° C. for 2 h. After the startingmaterial was consumed completely as detected by LCMS, the reactionsolution was added dropwise to water (20 mL) and stirred, and a solidwas precipitated. The resulting mixture was filtered to give P-III-30(60.2 mg, yield 61%) as a gray solid. MS-ESI: m/z 522.2 [M+H]+.

1H NMR (400 MHz, DMSO-d6) δ 8.42 (d, J=8.7 Hz, 1H), 7.79 (d, J=11.0 Hz,1H), 7.30 (s, 1H), 6.53 (s, 1H), 5.62-5.53 (m, 1H), 5.42 (s, 2H),5.30-5.16 (m, 2H), 4.63 (d, J=4.6 Hz, 1H), 4.09-3.99 (m, 1H), 3.22-3.11(m, 2H), 2.40 (s, 3H), 2.28 (dd, J=13.7, 7.2 Hz, 1H), 2.22-2.08 (m, 3H),1.94-1.78 (m, 2H), 1.08 (d, J=6.1 Hz, 3H), 0.87 (t, J=7.3 Hz, 3H).

Preparation Example 1.24.N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide

Step 1.

24a (19.6 mg, 0.188 mmol), KI4 (100 mg, 0.188 mmol) and DIEA (60.7 mg,0.47 mmol) were added to a three-necked flask (100 mL), and dissolvedwith DMF (2 mL). The solution was purged with N₂ three times, added withHATU (85.9 mg, 0.226 mmol) with stirring at 0° C. and reacted at 0° C.for 2 hours. After the reaction was completed as detected by TLC(DCM:MeOH=10:1), the reaction solution was added to water (20 mL), and agray solid was precipitated, lyophilized and purified on a preparationplate (EA:MeOH=10:1) to give P-III-31 (54.5 mg, yield 56%) as a paleyellow solid. MS-ESI: m/z 522.5 [M+H]+.

1H NMR (400 MHz, DMSO-d6) δ 8.44 (dd, J=8.7, 2.9 Hz, 1H), 7.78 (dd,J=10.9, 2.6 Hz, 1H), 7.30 (s, 1H), 6.53 (s, 1H), 5.60-5.51 (m, 1H), 5.42(s, 2H), 5.21 (dd, J=10.9, 3.4 Hz, 2H), 4.66 (dd, J=13.1, 4.7 Hz, 1H),4.10-3.98 (m, 1H), 3.21-3.10 (m, 2H), 2.39 (s, 3H), 2.34-2.05 (m, 4H),1.93-1.77 (m, 2H), 1.08 (dd, J=6.2, 1.6 Hz, 3H), 0.87 (t, J=7.3 Hz, 3H).

Preparation Example 1.25.(1R,4R)-4-(((S)-7-benzyl-20-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3,6,9,12,15-pentaoxo-2,5,8,11,14-pentaazaicosyl)oxy)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)cyclohexane-1-carboxamide

Step 1.

Benzyl bromide (8.90 g, 52.1 mmol) was added dropwise to a solution of25a (5.00 g, 34.7 mmol) and NaHCO₃ (8.74 g, 105 mmol) in DMF (50 mL)under nitrogen atmosphere, and the mixture was reacted at 25° C. for 17h. After the reaction was completed as detected by TLC (PE/EA=3/1), thereaction solution was added to water (250 mL), extracted with EA (150mL) separated, and washed with saturated aqueous sodium chloridesolution (200 mL). The organic phase was dried over anhydrous Na₂SO₄,concentrated and purified by column chromatography (PE:EA=7:1) to give acolorless liquid (4.60 g, yield 56.6%).

Step 2.

A solution of 25b (3.00 g, 1.00 mmol) in THF (5 mL) was added dropwiseto a solution of KI2 (2.36 g, 6.41 mmol) and TsOH (472 mg, 2.48 mmol) inTHF (15 mL) at 0° C. under nitrogen atmosphere, and the mixture wasreacted at 25° C. for 2 h. After the reaction was completed as detectedby TLC (PE/EA=1/1), the reaction solution was added to water (100 mL),extracted with EA (60 mL) twice and separated. The organic phase wasdried over anhydrous Na₂SO₄, concentrated and purified by columnchromatography (PE:EA=2:1) to give a white solid (4.00 g, yield 57.6%).

Step 3.

Pd/C (340 mg) was added to a mixed solution of 25c (1.70 g, 3.14 mmol)in MeOH (20 mL) and EA (20 mL) at 0° C. under hydrogen atmosphere, andthe mixture was reacted at 0° C. for 2 h. After the reaction wascompleted as detected by LCMS, the reaction solution was filteredthrough celite, and the filter cake was washed with EA (100 mL). Thefiltrate was concentrated, and washed with THF (10 mL) three times toremove MeOH to give a gray solid (530 mg, yield 21%).

Step 4.

DIEA (130 mg, 1.01 mmol) was added to a solution of 25d (200 mg, 0.443mmol), HY-13631A (214 mg, 0.402 mmol) and HATU (183 mg, 0.481 mmol) inDMF (4 mL) at 0° C. under nitrogen atmosphere, and the mixture wasreacted at 0° C. for 2 h. After the reaction was completed as detectedby LCMS, the reaction solution was added to a saturated aqueous citricacid solution (100 mL), and a brown solid was precipitated. Theresulting mixture was filtered, and the filter cake was washed withwater (100 mL) twice, dried by filtration, and dried with an oil pump togive a brown solid (300 mg, yield 78%).

Step 5.

DIEA (9 mL) was added dropwise to a solution of 25e (300 mg, 0.345 mmol)in DCM (40 mL) at 0° C. under nitrogen atmosphere, and the mixture wasreacted at 0° C. for 6 h. When there was 3% starting material left asdetected by LCMS, the reaction solution was added to a petroleumethersolution (600 mL) at 0° C., and a solid was precipitated. The resultingmixture was left to stand until the solid was adsorbed on the bottom ofthe flask, and the solution was poured out, washed with petroleumether(50 mL) twice, and dried with an oil pump to give a brown solid (178 mg,80%).

Step 6.

HATU (70.4 mg, 0.185 mmol) was added to a solution of 25f (80.0 mg,0.123 mmol), KI-1 (87.6 mg, 0.185 mmol) and DIEA (47.8 mg, 0.370 mmol)in DMF (2 mL) at 0° C. under nitrogen atmosphere, and the mixture wasreacted at 0° C. for 2 h. After the reaction was completed as detectedby LCMS, the reaction solution was added to an aqueous citric acidsolution (30 mL) at pH 4 at 0° C., and a flocculent solid wasprecipitated but could not be filtered off. The resulting solution wasextracted with DCM/MeOH (10/1, 100 mL) solution, separated and washedwith a saturated aqueous sodium chloride solution (50 mL). The organicphase was dried over anhydrous sodium sulfate, concentrated and purifiedon a preparation plate (DCM/MeOH=10/1) to give a pale yellow solid (17mg, yield 12.5%).

MS m/z (ESI): 1102 [M+1]

H-NMR (400 MHz, DMSO-D): 8.04 (m, 1H), 7.81 (d, 1H), 7.32 (s, 1H), 7.01(s, 2H), 6.54 (s, 1H), 5.61-5.51 (m, 1H), 5.48-5.40 (m, 2H), 5.18 (dd,2H), 4.65-4.59 (m, 4H), 3.80-3.68 (m, 8H), 3.23-3.17 (m, 2H), 2.42 (s,3H), 2.29-2.19 (m, 1H), 2.19-2.09 (m, 4H), 1.94-1.71 (m, 6H), 1.63-1.35(m, 10H), 1.25-1.18 (m, 2H), 0.89 (t, 3H)

Preparation Example 1.26.(1S,4S)-4-(((S)-7-benzyl-20-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3,6,9,12,15-pentaoxo-2,5,8,11,14-pentaazaicosyl)oxy)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)cyclohexane-1-carboxamide

Step 1

Benzyl bromide (8.90 g, 52.1 mmol) was added dropwise to a solution of26a (5.00 g, 34.7 mmol) and NaHCO₃ (8.74 g, 105 mmol) in DMF (50 mL)under nitrogen atmosphere, and the mixture was reacted at 25° C. for 17h. After the reaction was completed as detected by TLC (PE/EA=3/1), thereaction solution was added to water (250 mL), extracted with EA (150mL) separated and washed with saturated aqueous sodium chloride solution(200 mL). The organic phase was dried over anhydrous Na₂SO₄,concentrated and purified by column chromatography (PE:EA=7:1) to give acolorless liquid (4.60 g, yield 62.4%).

Step 2.

A solution of 26b (2.25 g, 9.62 mmol) in THF (5 mL) was added dropwiseto a solution of KI2 (1.77 g, 4.81 mmol) and TsOH (354 mg, 1.86 mmol) inTHF (20 mL) at 0° C. under nitrogen atmosphere, and the mixture wasreacted at 25° C. for 2 h. After the reaction was completed as detectedby TLC (PE/EA=1/1), the reaction solution was added to water (100 mL),extracted with EA (60 mL) twice and separated. The organic phase wasdried over anhydrous Na₂SO₄, concentrated and purified by columnchromatography (PE:EA=2:1) to give a white solid (1.2 g, yield 23%).

Step 3.

Pd/C (120 mg) was added to a mixed solution of 26c (600 mg, 3.14 mmol)in MeOH (15 mL) and EA (15 mL) at 0° C. under hydrogen atmosphere, andthe mixture was reacted at 0° C. for 2 h. After the reaction wascompleted as detected by LCMS, the reaction solution was filteredthrough celite, and the filter cake was washed with EA (50 mL). Thefiltrate was concentrated, and washed with THF (10 mL) three times toremove MeOH to give a gray solid (260 mg, yield 52%).

Step 4.

DIEA (121 mg, 0.940 mmol) was added to a solution of 26d (187 mg, 0.414mmol), HY-13631A (200 mg, 0.376 mmol) and HATU (171 mg, 0.451 mmol) inDME (4 mL) at 0° C. under nitrogen atmosphere, and the mixture wasreacted at 0° C. for 2 h. After the reaction was completed as detectedby LCMS, the reaction solution was added to a saturated aqueous citricacid solution (200 mL), and a brown solid was precipitated. Theresulting mixture was filtered, and the filter cake was washed withwater (100 mL) twice, dried by filtration, and dried with an oil pump togive a brown solid (230 mg, yield 70%).

Step 5.

Diethylamine (1.5 mL) was added dropwise to a solution of 26e (110 mg,0.126 mmol) in DCM (3 mL) at 0° C. under nitrogen atmosphere, and themixture was reacted at 0° C. for 6 h. When there was 3% startingmaterial left as detected by LCMS, the reaction solution was added to apetroleumether solution (500 mL) at 0° C., and a solid was precipitated.The resulting mixture was left to stand until the solid was adsorbed onthe bottom of the flask, and the solution was poured out, washed withpetroleumether (50 mL) twice, and dried with an oil pump to give a brownsolid (65 mg, 79%).

Step 6.

A solution of HATU (70.4 mg, 0.185 mmol) in DMF (1 mL) was added to asolution of 26f (60 mg, 0.093 mmol), KI-1 (66 mg, 0.14 mmol) and DIEA(36 mg, 0.28 mmol) in DMF (1 mL) at 0° C. under nitrogen atmosphere, andthe mixture was reacted at 0° C. for 2 h. After the reaction wascompleted as detected by LCMS, the reaction solution was added to anaqueous citric acid solution (30 mL) at pH 4 at 0° C., and a flocculentsolid was precipitated but could not be filtered off. The resultingsolution was extracted with DCM/MeOH (10/1, 100 mL) solution, separatedand washed with a saturated aqueous sodium chloride solution (50 mL).The organic phase was dried over anhydrous sodium sulfate, concentratedand purified on a preparation plate (DCM/MeOH=10/1) to give a paleyellow solid (12 mg, yield 11.7%).

MS m/z (ESI): 1102 [M+1]

H-NMR (400 MHz, MeOD): 7.70 (d, 1H), 7.66 (s, 1H), 7.35-7.20 (m, 5H),6.79 (s, 2H), 5.68-5.54 (m, 2H), 5.42-5.38 (m, 2H), 5.25 (dd, 2H),5.01-4.97 (m, 3H), 4.58-4.52 (m, 1H), 3.97-3.73 (m, 6H), 3.51-3.46 (m,3H), 3.28-3.01 (m, 3H), 2.47 (s, 3H), 2.31-2.26 (m, 4H), 2.20-2.12 (m,2H), 2.09-1.95 (m, 4H), 1.71-1.57 (m, 8H), 1.47-1.39 (m, 2H), 1.02 (t,3H)

Example 1.27

Step 1.

Benzyl bromide (11.0 g, 64.6 mmol) was added dropwise to a solution of27a (5.00 g, 43.0 mmol) and NaHCO₃ (10.9 g, 129 mmol) in DMF (50 mL)under nitrogen atmosphere, and the mixture was reacted at 25° C. for 17h. After the reaction was completed as detected by TLC (PE/EA=2/1), thereaction solution was added to water (500 mL), extracted with EA (250mL) twice, separated and washed with saturated aqueous sodium chloridesolution (500 mL). The organic phase was dried over anhydrous Na₂SO₄,concentrated and purified by column chromatography (PE:EA=3:2) to give acolorless liquid (5.1 g, yield 57.1%).

Step 2.

A solution of 27b (4.50 g, 21.8 mmol) in THF (10 mL) was added dropwiseto a solution of KI2 (4.00 g, 10.9 mmol) and TsOH (800 mg, 4.65 mmol) inTHF (30 mL) at 0° C. under nitrogen atmosphere, and the mixture wasreacted at 25° C. for 2 h. After the reaction was completed as detectedby TLC (PE/EA=1/2), the reaction solution was added to water (200 mL),extracted with EA (200 mL) twice and separated. The organic phase wasdried over anhydrous Na₂SO₄, concentrated and purified by columnchromatography (PE/EA=3/2) to give a white solid (1.56 g, yield 26%).

Step 3.

Pd/C (80 mg) was added to a mixed solution of 27c (800 mg, 1.55 mmol) inEtOH (8 mL) and EA (8 mL) at 0° C. under hydrogen atmosphere, and themixture was stirred at 0° C. for 2.5 h. After the reaction was completedas detected by LCMS, the reaction solution was filtered through celite,and the filter cake was washed with EA (200 mL). The filtrate wasconcentrated, dissolved with THF (20 mL) and dried by rotary evaporationto give a white solid (600 mg, yield 91%).

Step 4.

DIEA (152 mg, 1.18 mmol) was added to a solution of 27d (220 mg, 0.515mmol), HY-13631A (250 mg, 0.47 mmol) and HATU (214 mg, 0.56 mmol) in DMF(6 mL) at 0° C. under nitrogen atmosphere, and the mixture was reactedat 0° C. for 2 h. After the reaction was completed as detected by LCMS,the reaction solution was added to an aqueous citric acid solution(pH=4) (150 mL), and filtered. The filter cake was washed with water(175 mL), dried by filtration, and dried with an oil pump to give abrown solid (260 mg, yield 66%).

Step 5.

Diethylamine (8 mL) was added dropwise to a solution of 27e (260 mg,0.309 mmol) in DCM (30 mL) at 0° C. under nitrogen atmosphere, and themixture was reacted at 0° C. for 3 h. After the reaction was completedas detected by LCMS, the reaction solution was added to a petroleumethersolution (600 mL) at 0° C., and a solid was precipitated. The resultingmixture was left to stand until the solid was adsorbed on the bottom ofthe flask, and the solution was poured out and dried with an oil pump togive a brown solid (90 mg, yield 47.1%).

Step 6.

HATU (74 mg, 0.19 mmol) was added to a solution of 27f (90 mg, 0.13mmol), KI-1 (92 mg, 0.19 mmol) and DIEA (50 mg, 0.39 mmol) in DMF (2.5mL) at 0° C. under nitrogen atmosphere, and the mixture was reacted at0° C. for 2 h. After the reaction was completed as detected by LCMS, thereaction mixture was added to an aqueous citric acid solution (30 mL) atpH 4 at 0° C., and a flocculent solid was precipitated. The resultingmixture was filtered, and purified on a preparation plate(DCM/MeOH=10/1) to give a pale yellow solid (9.2 mg, yield 6%).

MS m/z (ESI): 1074 [M+1]

H-NMR (400 MHz, MeOD): 7.65 (d, 1H), 7.62 (s, 1H), 7.30-7.21 (m, 5H),6.79 (s, 2H), 5.69-5.65 (m, 1H), 5.57 (d, 1H), 5.43-5.10 (m, 3H), 4.70(d, 2H), 4.48-4.39 (m, 2H), 4.10-4.05 (m, 1H), 4.01-3.75 (m, 5H), 3.46(t, 2H), 3.22-3.15 (m, 2H), 3.07-3.00 (m, 1H), 2.75 (m, 1H), 2.62 (m,1H), 2.45 (s, 3H), 2.37-2.20 (m, 6H), 2.10-2.02 (m, 2H), 2.00-1.92 (m,2H) 1.68-1.57 (m, 6H), 1.01 (t, 3H)

Example 1.28

Step 1.

Benzyl bromide (5.52 g, 32.0 mmol) was added dropwise to a solution of28a (2.50 g, 21.0 mmol) and NaHCO₃ (5.43 g, 64.0 mmol) in DMF (25 mL)under nitrogen atmosphere, and the mixture was reacted at 25° C. for 12h. After the reaction was completed as detected by TLC (PE/EA=2/1), thereaction solution was added to water (250 mL), extracted with EA (500mL), separated and washed with saturated aqueous sodium chloridesolution (500 mL). The organic phase was dried over anhydrous Na₂SO₄,concentrated and purified by column chromatography (PE:EA=3:2) to give acolorless liquid (2.50 g, yield 56.3%).

Step 2.

A solution of 28b (2.5 g, 12.1 mmol) in THF (1 mL) was added dropwise toa solution of KI2 (2.23 g, 6.05 mmol) and TsOH (446 mg, 2.59 mmol) inTHF (19 mL) at 0° C. under nitrogen atmosphere, and the mixture wasreacted at 25° C. for 2 h. After the reaction was completed as detectedby LCMS, the reaction solution was added to water (50 mL), extractedwith DCM (20 mL) three times and separated. The organic phase was driedover anhydrous Na₂SO₄, concentrated and purified by columnchromatography (PE/EA=3/2) to give a white solid (1.60 g, yield 43%).

Step 3.

Pd/C (80 mg) was added to a mixed solution of 28c (800 mg, 1.56 mmol) inMeOH (8 mL) and EA (8 mL) at 0° C. under hydrogen atmosphere, and themixture was stirred at 0° C. for 7 h. After the reaction was completedas detected by LCMS, the reaction solution was filtered through celite,and the filter cake was washed with EA (200 mL). The filtrate wasconcentrated to give a white solid (600 mg, yield 91%).

Step 4.

DIEA (122 mg, 0.940 mmol) was added to a solution of 28d (176 mg, 0.414mmol), HY-13631A (200 mg, 0.377 mmol) and HATU (172 mg, 0.450 mmol) inDMF (7 mL) at 0° C. under nitrogen atmosphere, and the mixture wasreacted at 0° C. for 2 h. After the reaction was completed as detectedby LCMS, the reaction solution was added to a saturated aqueous citricacid solution (120 mL). The resulting mixture was filtered, and thefilter cake was washed with water (120 mL) once, dried by filtration,and dried with an oil pump to give a brown solid (280 mg, yield 88%).

Step 5.

DIEA (4.2 mL) was added dropwise to a solution of 28e (140 mg, 0.166mmol) in DCM (14 mL) at 0° C. under nitrogen atmosphere, and the mixturewas reacted at 0° C. for 7 h. When there was 2% starting material leftas detected by LCMS, the reaction solution was added to a petroleumethersolution (420 mL) at 0° C., and a solid was precipitated. The resultingmixture was left to stand until the solid was adsorbed on the bottom ofthe flask, and the solution was poured out, washed with petroleumether(50 mL) three times, and dried with an oil pump to give a brown solid(97 mg, yield 68%).

Step 6.

HATU (91.2 mg, 0.240 mmol) was added to a solution of 28f (82.0 mg,0.130 mmol), KI-1 (113 mg, 0.240 mmol) and DIEA (51 mg, 0.39 mmol) inDMF (5 mL) at 0° C. under nitrogen atmosphere, and the mixture wasreacted at 0° C. for 2 h. After the reaction was completed as detectedby LCMS, the reaction mixture was added to an aqueous citric acidsolution (30 mL) at pH 4 at 0° C., and a flocculent solid wasprecipitated. The resulting mixture was filtered, and purified on apreparation plate (DCM/MeOH=10/1) to give a pale yellow solid (24 mg,yield 17%).

MS m/z (ESI): 1074 [M+1]

H-NMR (400 MHz, MeOD): 7.66 (d, 1H), 7.62 (s, 1H), 7.27-7.20 (m, 5H),6.76 (s, 2H), 5.70-5.62 (m, 1H), 5.58 (d, 1H), 5.38-5.17 (m, 3H), 4.73(d, 1H), 4.62-4.52 (m, 2H), 4.50-4.39 (m, 2H), 3.90-3.83 (d, 4H),3.72-3.67 (d, 1H), 3.49-3.41 (m, 1H), 3.25-3.11 (m, 3H), 3.07-2.98 (m,2H), 2.61-2.52 (m, 2H), 2.43 (s, 3H), 2.31-2.20 (m, 6H), 2.01-1.90 (m,2H), 2.00-1.92 (m, 2H), 1.65-1.52 (m, 4H), 1.30-1.25 (m, 2H), 1.00 (t,3H)

Preparation Example 1.29. N-((7S)-7-benzyl-1-((14(1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-2-oxopyrrolidin-3-yl)oxy)-3,6,9,12-tetraoxo-2,5,8,11-tetraazatridecan-13-yl)-6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamide

Step 1.

KI2 (638 mg, 1.73 mmol) and TsOH (50 mg, 0.29 mmol) were added to asolution of 29a (300 mg, 0.577 mmol) in DMA (30 mL) sequentially, andthe mixture was stirred at 50° C. for 3 h. The reaction solution wascooled to room temperature, supplemented with KI2 (1.28 g, 3.46 mmol)and stirred at 50° C. for 17 h. When there was about 50% productgenerated as detected by LCMS, the reaction solution was cooled to roomtemperature, added to ice water, extracted with DCM/MeOH (5:1, 30 mL)three times, washed with a saturated aqueous sodium chloride solution(50 mL), and separated. The organic phase was dried over anhydroussodium sulfate, concentrated, and purified by column chromatography togive an impure product, which was separated by prep-HPLC (10 mmol/LNH₄OAc) to give a white solid (70 mg, yield 12%).

Step 2.

Diethylamine (3.0 mL) was added to a solution of 29b (70 mg, 0.085 mmol)in DCM (9 mL) at 0° C., and the mixture was stirred at 0° C. for 9 h.After the reaction was completed as detected by LCMS, the reactionsolution was added to petroleumether (250 mL) at 0° C., and a solid wasprecipitated. The mixture was stirred for 10 min, and petroleumether waspoured out. A three-necked flask was washed with petroleumether (50 mL)and dried with a pump, and the product was transferred to asingle-necked flask with tetrahydrofuran (20 mL) and dried by rotaryevaporation to give a yellow solid (40 mg, yield 78%).

Step 3.

HATU (45 mg, 0.012 mmol) and DIEA (26 mg, 0.20 mmol) were added to asolution of 29c (40 mg, 0.066 mmol) and Kul (56 mg, 0.012 mmol) in DME(2 mL) at 0° C., and the mixture was stirred for 1 h. After the reactionwas completed as detected by LCMS, the reaction solution was poured intoa citric acid solution (20 mL) at pH 4-5, extracted with DCM/MeOH (5/1,30 mL), and washed with saturated brine (10 mL). The organic phase wasdried over anhydrous sodium sulfate, concentrated, and purified byprep-TLC (DCM:MeOH=10:1) to give a yellow solid (15 mg, yield 21%).

MS m/z (ESI): 1060 [M+1]

H-NMR (400 MHz, DMSO-D): 7.81 (d, 1H), 7.32 (s, 1H), 7.24 (m, 5H), 7.00(s, 2H), 6.55 (s, 1H), 5.45 (dd, 2H), 5.11-4.80 (m, 4H), 4.60-4.50 (m,1H), 4.40-4.30 (m, 1H), 3.80-3.71 (m, 3H), 3.70-3.66 (m, 2H), 3.65-3.60(m, 1H), 3.30-2.79 (m, 6H), 2.46-2.35 (m, 6H), 2.25-2.17 (m, 3H),1.94-1.80 (m, 3H), 1.50-1.40 (m, 4H), 1.28-1.20 (m, 2H), 1.00 (t, 3H)

Preparation Example 1.30.N—((S)-7-benzyl-17-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)amino)-2,5,8,11,17-pentaoxo-14-thia-3,6,9,12-tetraazaheptadecyl)-6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamide

Step 1.

TsOH (52 mg, 0.272 mmol) and P12-3 (288 mg, 2.72 mmol) were added to asolution of 30a (1.00 g, 2.72 mmol) in THF (10 mL) at 0° C. undernitrogen atmosphere. After the addition was completed, the mixture wasslowly warmed to 25° C. and reacted for 1 h. The reaction solution wasdirectly mixed with silica gel and purified by column chromatography(DCM/MeOH=80/1) to give a white solid (760 mg, yield 67%).

Step 2.

DIEA (181 mg, 1.40 mmol) was added to a solution of 30b (321 mg, 0.620mmol), HY-13631A (300 mg, 0.560 mmol) and HATU (255 mg, 0.670 mmol) inDMF (12 mL) under nitrogen atmosphere. After the addition was completed,the mixture was reacted at 25° C. for 2 h. After the reaction wascompleted as detected by TLC (DCM/MeOH=20/1), the reaction solution wasadded dropwise to water (180 mL), filtered and purified on a preparationplate (DCM/MeOH=20/1) to give a yellow solid (320 mg, yield 68%).

Step 3.

Diethylamine (4 mL) was added dropwise to a solution of 30c (115 mg,0.138 mmol) in DCM (12 mL) under nitrogen atmosphere, and the mixturewas stirred at 0° C. for 3 h. After the starting material was consumedcompletely as detected by LCMS, the reaction solution was added dropwiseinto PE (345 mL) at 0° C. under nitrogen atmosphere, and stirred for 3min. The resulting mixture was left to stand at 0° C. for 1 h until thesolid was gathered at the bottom of the flask, and the supernatant waspoured out, and dried with a pump to give a yellow solid (56 mg, yield67%).

Step 4.

HATU (57 mg, 0.15 mmol) was added to a solution of 30d (51 mg, 0.084mmol), Kul (71.1 mg, 0.151 mmol) and DIEA (33 mg, 0.25 mmol) in DMF (2.5mL) at 0° C. under nitrogen atmosphere, and the mixture was reacted at0° C. for 2 h. After the starting material was consumed completely asdetected by LCMS, the reaction solution was added dropwise into citricacid (30 mL) at pH 4 at 0° C.

The resulting mixture was filtered to give a yellow solid (130 mg),which was purified by prep-HPLC (10 mmol/L NH₄OAc) to give a white solid(22 mg, yield 25%).

MS m/z (ESI): 1064 [M+1]

H-NMR (400 MHz, MeOD): 7.70 (d, 1H), 7.65 (s, 1H), 7.21 (m, 3H), 7.07(d, 2H), 6.80 (s, 1H), 5.70 (m, 1H), 5.47 (dd, 4H), 4.66 (m, 2H), 4.33(m, 3H), 3.82 (m, 5H), 3.61 (m, 1H), 3.52 (m, 3H), 3.25 (m, 1H),3.06-2.75 (m, 4H), 2.62 (m, 2H), 2.47 (s, 3H), 2.30 (m, 2H), 2.25 (m,2H), 1.60 (m, 4H), 1.25 (m, 2H), 0.99 (t, 3H)

Preparation Example 1.31.N-((7S,15R)-7-benzyl-17-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamide

Step 1.

Benzyl bromide (24.6 g, 144 mmol) was added dropwise to a solution of31a (10.0 g, 96.0 mmol) and NaHCO₃ (24.1 g, 288 mmol) in DMF (50 mL)under nitrogen atmosphere, and the mixture was reacted at 25° C. for 17h. After the reaction was completed as detected by TLC (PE/EA=1/1), thereaction solution was added to water (500 mL), extracted with EA (200mL) twice, separated and washed with saturated aqueous sodium chloridesolution (200 mL). The organic phase was dried over anhydrous Na₂SO₄,concentrated and purified by column chromatography (PE:EA=3:2) to give apale yellow oily liquid (9.5 g, yield 57.1%).

Step 2.

A solution of 31b (6.30 g, 32.5 mmol) in THF (10 mL) was added dropwiseto a solution of KI2 (4.00 g, 10.9 mmol) and TsOH (800 mg, 4.65 mmol) inTHF (30 mL) at 0° C. under nitrogen atmosphere, and the mixture wasreacted at 25° C. for 2 h. After the reaction was completed as detectedby TLC (PE/EA=1/1), the reaction solution was added to water (400 mL),extracted with EA (200 mL) twice, separated and washed with saturatedaqueous sodium chloride solution (200 mL) once. The organic phase wasdried over anhydrous Na₂SO₄, concentrated and purified by columnchromatography (PE/EA=3/2) to give a white sticky solid (850 mg, yield15.5%).

Step 3.

Pd/C (200 mg) was added to a mixed solution of 31c (500 mg, 0.990 mmol)in EtOH (5 mL) and EA (5 mL) at 0° C. under hydrogen atmosphere, and themixture was reacted at 0° C. for 2.5 h. After the reaction was completedas detected by LCMS, the reaction solution was filtered through celite,and the filter cake was washed with EA (200 mL). The filtrate wasconcentrated, and dissolved with THF (20 mL) and dried by rotaryevaporation, which was repeated three times, to give a gray solid (350mg, yield 85%).

Step 4.

DIEA (152 mg, 1.18 mmol) was added to a solution of 31d (213 mg, 0.515mmol), HY-13631A (250 mg, 0.470 mmol) and HATU (214 mg, 0.560 mmol) inDMF (6 mL) at 0° C. under nitrogen atmosphere, and the mixture wasreacted at 0° C. for 2 h. After the reaction was completed as detectedby LCMS, the reaction solution was added to a saturated aqueous citricacid solution (150 mL), and a brown solid was precipitated. Theresulting mixture was filtered, and the filter cake was washed withwater (175 mL), dried by filtration, and dried with an oil pump to givea brown solid (150 mg, yield 38.5%).

Step 5.

Diethylamine (10 mL) was added dropwise to a solution of 31e (100 mg,0.120 mmol) in DCM (30 mL) at 0° C. under nitrogen atmosphere, and themixture was reacted at 0° C. for 6 h. After the reaction was completedas detected by LCMS, the reaction solution was added to a petroleumethersolution (600 mL) at 0° C., and a solid was precipitated. The resultingmixture was left to stand until the solid was adsorbed on the bottom ofthe flask, and the solution was poured out and dried with an oil pump togive a gray solid (95 mg, yield not counted, impure product).

Step 6.

A solution of HATU (74 mg, 0.19 mmol) in DMF (1 mL) was added to asolution of 31f (90 mg, 0.15 mmol), Kul (140 mg, 0.296 mmol) and DIEA(57 mg, 0.44 mmol) in DMF (2 mL) at 0° C. under nitrogen atmosphere, andthe mixture was reacted at 0° C. for 2 h. After the reaction wascompleted as detected by LCMS, the reaction mixture was added to anaqueous citric acid solution (30 mL) at pH 4 at 0° C., and a flocculentsolid was precipitated. The resulting mixture was filtered, and purifiedby prep-TLC (DCM/MeOH=10/1) to give a pale yellow solid (18.7 mg, yield3% over two steps).

MS m/z (ESI): 1062 [M+1]

H-NMR (400 MHz, MeOD): 7.65 (m, 2H), 7.21 (m, 5H), 6.80 (s, 1H), 5.60(m, 2H), 5.41 (m, 2H), 5.19 (m, 1H), 4.72 (s, 2H), 4.47 (m, 1H), 4.25(m, 1H), 3.84 (m, 5H), 3.65 (m, 1H), 3.48 (m, 2H), 3.06 (m, 1H), 2.95(m, 1H), 2.50 (m, 5H), 2.27 (m, 5H), 1.98 (m, 2H), 1.60 (m, 5H), 1.33(m, 2H), 1.03 (t, 3H)

Preparation Example 1.32.N-((7S,15S)-7-benzyl-17-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamide

Step 1.

Bromopropene (960 mg, 7.92 mmol) was added to a solution of 32a (2.00 g,6.6 mmol) and

K₂CO₃ (1.82 g, 13.2 mmol) in MeCN (20 mL), and the mixture was stirredat 20° C. for 5 h. After the reaction was completed as detected by TLC(PE/EA=1/2), the reaction solution was poured into water (100 mL),adjusted to pH 5 and extracted with EA (100 mL) three times. The organicphase was dried over anhydrous sodium sulfate, dried by rotaryevaporation, and purified by column chromatography (PE/EA=2/1) to give32b (1.83 g, yield 81%) as a white solid.

Step 2.

TFA (10 mL) was added to a solution of 32b (1.38 g, 4.02 mmol) in DCM(10 mL), and the mixture was stirred at 25° C. for 17 h. After thereaction was completed as detected by TLC (PE/EA=1/3), the reactionsolution was dried by rotary evaporation to give 32c (0.91 g, yield notcounted) as a yellow sticky substance.

Step 3.

41d (1.92 g, 4.87 mmol) was added to a solution of 32c (910 mg, 4.87mmol) and NaHCO₃ (613 mg, 7.3 mmol) in DME/H₂O (20 mL/10 mL), and themixture was stirred at 25° C. for 3 h. After the reaction was completedas detected by TLC (DCM/MeOH=1/1), the reaction solution was poured intowater (100 mL), adjusted to pH 5 with aq. HCl (1 N) and extracted withEA (150 mL) twice. The organic phase was dried over anhydrous sodiumsulfate, dried by rotary evaporation, and purified by columnchromatography (DCM/MeOH=20/1) to give 32e (1.53 g, yield 67%) as awhite solid. MS-ESI: m/z 467.4 [M+H]+.

Step 4.

Pd/C (600 mg) was added to a solution of 32f (3 g, 5.83 mmol) in MeOH(50 mL), and the mixture was stirred under hydrogen atmosphere at 25° C.for 5 h. After the reaction was completed as detected by TLC (EA), thereaction solution was filtered and dried by rotary evaporation to give32 g (1.9 g, yield 77%) as a white solid.

Step 5.

HATU (707 mg, 1.86 mmol) was added to a solution of 32 g (789 mg, 1.86mmol), KI4 (900 mg, 1.69 mmol) and triethylamine (342 mg, 3.38 mmol) inDMF (10 mL), and the mixture was stirred at 0° C. for 3.5 h. After thereaction was completed as detected by TLC (EA), the reaction solutionwas poured into H₂O (80 mL), and extracted with EA (100 mL) twice. Theorganic phase was dried over anhydrous sodium sulfate, dried by rotaryevaporation, and purified by column chromatography (EA) to give 32 h(1.186 g, yield 83%) as a white solid. MS-ESI: m/z 842.3 [M+H]+.

Step 6.

A solution of 32 h (1.186 g, 1.41 mmol) in DCM/diethylamine (20 mL,20/1) was stirred at 25° C. for 17 h. After the reaction was completedas detected by TLC (DCM/MeOH=10/1), the reaction solution was pouredinto petroleumether (200 mL), and the resulting mixture was filtered togive 32i (768 mg, yield 88%) as a white solid. MS-ESI: m/z 620.3 [M+H]+.

Step 7.

HATU (414 mg, 1.09 mmol) was added to a solution of 32 g (676 mg, 1.09mmol), 41e (508 mg, 1.09 mmol) and DIEA (423 mg, 3.27 mmol) in DMF (10mL), and the mixture was stirred at 20° C. for 17 h. After the reactionwas completed as detected by TLC (PE/EA=1/5), the reaction solution waspoured into water (30 mL). The resulting mixture was filtered, and thefilter cake was purified by column chromatography (DCM/MeOH=50/1) togive 32j (511 mg, yield 44%) as a white solid. MS-ESI: m/z 1068.3[M+H]+.

Step 8.

A solution of 32j (482 mg, 0.451 mmol) in diethylamine/DCM (10 mL, 1/5)was stirred at 10° C. for 17 h. After the reaction was completed asdetected by TLC (EA), the reaction solution was poured into PE (300 mL),and the resulting mixture was filtered to give 32k (301 mg, yield notcounted) as a white solid.

Step 9.

Morpholine (93 mg, 1.07 mmol) was added to a solution of 32k (301 mg,0.356 mmol) and Pd(PPh₃)₄ (82 mg, 0.071 mmol) in THF (5 mL), and themixture was stirred at 25° C. for 5 h. After the reaction was completedas detected by LCMS, the reaction solution was purified by preparativechromatography to give 321 (108 mg, yield 38%) as a white solid. MS-ESI:m/z 806.3 [M+H]+.

Step 10.

Bromoacetyl bromide (27 mg, 0.134 mmol) was added to a solution of 321(108 mg, 0.134 mmol) and triethylamine (41 mg, 0.402 mmol) in THF (2 mL)and DMF (2 mL), and the mixture was stirred at 0° C. for 1 h. After thereaction was completed as detected by TLC (DCM/MeOH=10/1), the reactionsolution was directly purified by preparative chromatography to giveL-II-27 (15 mg, yield 12%) as a white solid.

MS-ESI: m/z 926.3 [M+H]+.

1H NMR (400 MHz, DMSO-d6) δ 12.11 (s, 1H), 8.54-8.42 (m, 3H), 8.27-8.16(m, 2H), 7.78 (d, J=11.0 Hz, 1H), 7.30 (s, 1H), 6.53 (s, 1H), 5.61-5.51(m, 1H), 5.42 (s, 2H), 5.20-5.05 (m, 2H), 4.56-4.42 (m, 2H), 4.32-4.22(m, 1H), 3.96-3.87 (m, 3H), 3.79 (d, J=5.6 Hz, 2H), 3.70 (d, J=5.9 Hz,2H), 3.25-3.08 (m, 2H), 2.61-2.53 (m, 2H), 2.45-2.36 (m, 4H), 2.36-2.22(m, 3H), 2.20-2.03 (m, 4H), 1.99-1.68 (m, 4H), 0.87 (t, J=7.3 Hz, 3H).

Preparation Example 1.33(1R,3R)-3-(((S)-7-benzyl-16-bromo-3,6,9,12,15-pentaoxo-2,5,8,11,14-pentaazahexadecyl)oxy)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)cyclobutane-1-carboxamide

Step 1.

Pd/C (400 mg, 10 wt. %) was added to a solution of 33a (2.00 g, 2.58mmol) in MeOH (20 mL), and the mixture was stirred at 20° C. for 5 h.After the reaction was completed as detected by TLC (EA), the reactionsolution was filtered and dried by rotary evaporation to give 33b (1.3g, yield 74%) as a white solid.

Step 2.

HATU (305 mg, 0.802 mmol) was added to a solution of 33b (0.55 g, 0.802mmol), KI4 (427 mg, 0.802 mmol) and DIPEA (310 mg, 2.40 mmol) in DMF (5mL), and the mixture was stirred at 0° C. for 2 h. After the reactionwas completed as detected by TLC (DCM/MeOH=1/10), the reaction solutionwas poured into water (40 mL). The resulting mixture was filtered, andthe filter cake was purified by column chromatography (DCM/MeOH=20/1) togive 33c (360 mg, yield 41%) as a yellow solid.

Step 3.

Diethylamine (2 mL) was added to a solution of 33c (360 mg, 0.326 mmol)in DCM (10 mL), and the mixture was stirred at 25° C. for 17 h. Afterthe reaction was completed as detected by TLC (DCM/MeOH=5/1), thereaction solution was poured into PE (100 mL), and the resulting mixturewas filtered to give 33d (205 mg, yield 71%) as a white solid. MS-ESI:m/z 881.3 [M+H]+.

Step 4.

A solution of bromoacetyl bromide (94 mg, 0.446 mmol) in THF (2 mL) wasadded to a solution of 33d (205 mg, 0.233 mmol) and triethylamine (118mg, 1.17 mmol) in DMF (1 mL) and water (1 mL), and the mixture wasstirred at 0° C. for 1 h. The reaction solution was purified bypreparative chromatography to give L-II-28 (15 mg, yield 6%) as a whitesolid.

MS-ESI: m/z 1001.2 [M+H]+.

1H NMR (400 MHz, DMSO-d6) δ 8.57-8.50 (m, 1H), 8.50-8.43 (m, 2H),8.35-8.29 (m, 1H), 8.19-8.12 (m, 2H), 7.80 (d, J=10.8 Hz, 1H), 7.27-7.14(m, 7H), 6.53 (s, 1H), 5.59-5.51 (m, 1H), 5.44-5.39 (m, 2H), 5.20-5.07(m, 2H), 4.56-4.44 (m, 3H), 3.92 (s, 3H), 3.80-3.68 (m, 5H), 3.41 (s,1H), 3.21-3.12 (m, 2H), 2.83-2.74 (m, 1H), 2.58-2.55 (m, 3H), 2.39 (s,4H), 2.18-2.03 (m, 4H), 1.93-1.78 (m, 2H), 0.87 (t, J=7.3 Hz, 3H).

Preparation Example 1.34.N₄₇S,15S)-7-benzyl-17-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamide

Step 1.

34a (5 g, 48.0 mmol) and K₂CO₃ (19.9 g, 144.0 mmol) were dissolved inDMF (20 mL), followed by the dropwise addition of benzyl bromide (12.3g, 72.0 mmol). The mixture was reacted at 25° C. for 17 h. After thestarting material was consumed completely as detected by TLC(PE/EA=3/1), the reaction solution was added to water (200 mL),extracted with EA (250 mL), separated and washed with saturated NaCl.The organic phase was dried over anhydrous Na₂SO₄, concentrated andpurified by column chromatography (PE:EA=2:1) to give 34b (8.7 g, yield93%) as a colorless liquid. MS-ESI: m/z 195.1 [M+H]+.

Step 2.

34c (7.3 g, 19.8 mmol) and TsOH (1.46 g, 8.5 mmol) were dissolved in THF(20 mL), and the mixture was cooled to 0° C. under nitrogen atmosphere,added dropwise with a solution of 43b (7.7 g, 39.6 mmol) in THF (10 mL)and reacted at 0° C. for 2 h after the addition was completed. Aftermost of the starting material was consumed as detected by TLC(PE/EA=2/1), the reaction solution was poured into water (100 mL),extracted with DCM (100 mL), separated and washed with saturated NaCl.The organic phase was dried over anhydrous Na₂SO₄ and purified by columnchromatography (PE/EA=1/1) to give 34d (3.9 g, yield 39%) as a colorlesssticky substance. MS-ESI: m/z 503.3 [M+H]+.

Step 3.

Pd/C (1 g, 10 wt. %) was added to a mixed solution of 34d (1.9 g, 3.78mmol) in EtOH (100 mL) and EA (100 mL) at 0° C. under hydrogenatmosphere, and the mixture was reacted at 0° C. for 3 h. After thereaction was completed as detected by TLC (PE/EA=2/1), the reactionsolution was filtered through celite, and the filter cake was washedwith EA/EtOH (1:1, 100 mL×3). The filtrate was concentrated, anddissolved with THF (50 mL×3) and dried by rotary evaporation, which wasrepeated three times, to give 34e (1 g, yield 64%) as a gray solid.MS-ESI: m/z 435.2 [M+H]+.

Step 4.

DIEA (303 mg, 2.35 mmol) was added dropwise to a solution of 34e (426mg, 1.03 mmol), KI4 (500 mg, 0.94 mmol) and HATU (429 mg, 1.13 mmol) inDMF (20 mL) at 0° C. under nitrogen atmosphere, and the mixture wasreacted at 0° C. for 2 h after the addition was completed. After thereaction was completed as detected by LCMS, the reaction solution wasadded dropwise to water (300 mL) and stirred. The resulting mixture wasleft to stand for 5 min and filtered, and the filter cake was dissolvedwith DCM/MeOH (10:1, 100 mL) solution, dried by rotary evaporation,mixed with silica gel and purified by column chromatography(EA:MeOH=30:1) to give 34f (600 mg, yield 77%) as a yellow solid.MS-ESI: m/z 830.3 [M+H]+.

Step 5.

Diethylamine (5 mL) was added dropwise to a solution of 34f (150 mg,0.18 mmol) in DCM (5 mL) at 0° C. under nitrogen atmosphere, and themixture was reacted at 0° C. for 2 h. After the reaction was completedas detected by LCMS, a petroleumether solution (100 mL×6) was added tothe reaction solution, and a solid was precipitated. The resultingmixture was left to stand until the solid was adsorbed on the bottom ofthe flask, and the solution was poured out and dried with an oil pump togive 34 g (120 mg, yield 76%) as a white powder, with the productcontent of 70% as detected by LCMS. MS-ESI: m/z 608.3 [M+H]+.

Step 6.

A solution of HATU (45 mg, 0.118 mmol) in DMF (1 mL) was added to asolution of 34 g (60 mg, 0.099 mmol), 43 h (51 mg, 0.108 mmol) and DIEA(32 mg, 0.25 mmol) in DMF (1 mL) at 0° C. under nitrogen atmosphere, andthe mixture was reacted at 0° C. for 2 h. After the starting materialwas consumed completely as detected by LCMS, the reaction solution wasdirectly purified by reverse-phase column chromatography (eluent:(MeCN/MeOH=1/1):H₂O=60%:40%) to give L-III-30 (14.8 mg, yield 14%) as ayellow solid.

MS-ESI: m/z 1062.4 [M+H]+.

1H NMR (400 MHz, Methanol-d4) δ 7.69-7.61 (m, 2H), 7.22-7.16 (m, 2H),7.16-7.09 (m, 3H), 6.76 (s, 2H), 5.70-5.64 (m, 1H), 5.60 (d, J=16.4 Hz,1H), 5.40-5.31 (m, 2H), 5.26 (d, J=19.0 Hz, 1H), 4.65-4.50 (m, 7H),4.25-4.16 (m, 1H), 3.87 (d, J=16.7 Hz, 1H), 3.83-3.76 (m, 3H), 3.72 (d,J=17.0 Hz, 2H), 3.44 (t, J=7.1 Hz, 2H), 3.25-3.17 (m, 2H), 3.10-3.02 (m,1H), 2.92-2.83 (m, 1H), 2.45-2.39 (m, 5H), 2.32-2.20 (m, 5H), 1.97-1.89(m, 2H), 1.63-1.50 (m, 4H), 1.34-1.20 (m, 6H), 0.99 (t, J=7.3 Hz, 3H).

Preparation Example 1.35.N-((7S)-7-benzyl-17-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamide

Step 1.

35a (10.0 g, 96 mmol) and K₂CO₃ (39.8 g, 288 mmol) were dissolved in DMF(100 mL), followed by the dropwise addition of benzyl bromide (24.6 g,144 mmol). The mixture was reacted at room temperature for 17 h. Afterthe reaction was completed as detected by TLC (PE/EA=3/1), the reactionsolution was added to water (200 mL), extracted with EA (250 mL),separated and washed with saturated NaCl. The organic phase was driedover anhydrous Na₂SO₄, concentrated and purified by columnchromatography (PE:EA=4:1) to give 35b (15 g, yield 80%) as a colorlessliquid. MS-ESI: m/z 195.1 [M+H]+.

Step 2.

35c (6.6 g, 18 mmol) and 35b (7 g, 36 mmol) were dissolved in THF (50mL), and the mixture was cooled to 0° C. under nitrogen atmosphere,added dropwise with a solution of TsOH (1.3 g, 7.5 mmol) in THF (10 mL)and reacted at 0° C. for 2 h after the addition was completed. Aftermost of the starting material was consumed as detected by TLC(PE/EA=2/1), the reaction solution was poured into water (100 mL),extracted with DCM (100 mL), separated and washed with saturated NaCl.The organic phase was dried over anhydrous Na₂SO₄ and purified by columnchromatography (PE/EA=1/1) to give 35d (4.9 g, yield 54%) as a paleyellow sticky substance. MS-ESI: m/z 503.2 [M+H]+.

Step 3.

Pd/C (2 g, 10 wt. %) was added to a mixed solution of 35d (2.2 g, 4.4mmol) in EtOH (20 mL) and EA (20 mL) at 0° C. under hydrogen atmosphere,and the mixture was reacted at 0° C. for 3 h. After the reaction wascompleted as detected by TLC (PE/EA=2/1), the reaction solution wasfiltered through celite, and the filter cake was washed with EA/EtOH(1:1, 100 mL×3). The filtrate was concentrated, and dissolved with THF(50 mL×3) and dried by rotary evaporation, which was repeated threetimes, to give 35e (1.5 g, yield 83%) as a white solid. MS-ESI: m/z435.1 [M+H]+.

Step 4.

DIEA (606 mg, 4.70 mmol) was added dropwise to a solution of 35e (852mg, 2.07 mmol), KI4 (1 g, 1.88 mmol) and HATU (856 mg, 2.25 mmol) in DMF(50 mL) at 0° C. under nitrogen atmosphere, and the mixture was reactedat 0° C. for 2 h after the addition was completed. After the reactionwas completed as detected by LCMS, the reaction solution was addeddropwise to water (700 mL) and stirred. The resulting mixture was leftto stand for 5 min and filtered, and the filter cake was dissolved withDCM/MeOH (10:1, 150 mL) solution, dried by rotary evaporation, mixedwith silica gel and purified by column chromatography (EA:MeOH=30:1) togive 35f (900 mg, yield 79%) as a yellow solid. MS-ESI: m/z 830.3[M+H]+.

Step 5.

Diethylamine (1.5 mL) was added dropwise to a solution of 35f (800 mg,0.96 mmol) in DCM (30 mL) at 0° C. under nitrogen atmosphere, and themixture was reacted at RT for 17 h. After the reaction was completed asdetected by LCMS, a petroleumether solution (100 mL×6) was added to thereaction solution, and a solid was precipitated. The resulting mixturewas left to stand until the solid was adsorbed on the bottom of theflask, and the solution was poured out and dried with an oil pump togive a white powder (640 mg). Then the solid was treated with TFA:MeOH(1:20) to give 35 g (770 mg, yield 92%) as a yellow solid, with thecontent of 70% as detected by LCMS. MS-ESI: m/z 608.3 [M+H]+.

Step 6.

A solution of HATU (487 mg, 1.28 mmol) in DMF (2 mL) was added to asolution of 35 g (770 mg, 0.89 mmol), 43 h (480 mg, 1.02 mmol) and DIEA(345 mg, 2.67 mmol) in DMF (8 mL) under nitrogen atmosphere, and themixture was reacted at 0° C. for 2 h. After the starting material wasconsumed completely as detected by LCMS, the reaction solution was addedto ice water (100 mL) and stirred, and a yellow solid was precipitated.The resulting mixture was left to stand until the solid was absorbed thebottom of the flask and to the solution, the solution was frozen toprecipitate the product and filtered, and the solid was lyophilized,mixed with silica gel, and purified by column chromatography(EA/MeOH=20/1) to give L-III-31 (568 mg, yield 42%) as a pale yellowsolid.

MS-ESI: m/z 1062.8 [M+H]+.

1H NMR (400 MHz, DMSO-d6) δ 8.52-8.44 (m, 2H), 8.33-8.26 (m, 1H),8.15-8.09 (m, 1H), 8.09-8.04 (m, 1H), 8.03-7.96 (m, 1H), 7.81 (d, J=10.9Hz, 1H), 7.33 (s, 1H), 7.26-7.11 (m, 5H), 6.99 (s, 2H), 6.54 (s, 1H),5.61-5.52 (m, 1H), 5.46-5.40 (m, 2H), 5.28 (d, J=19.1 Hz, 1H), 5.17 (d,J=18.9 Hz, 1H), 4.69-4.59 (m, 1H), 4.59-4.45 (m, 2H), 4.08-3.98 (m, 1H),3.78-3.54 (m, 6H), 3.38-3.35 (m, 2H), 3.22-3.09 (m, 2H), 3.07-2.98 (m,1H), 2.82-2.72 (m, 1H), 2.43-2.35 (m, 4H), 2.27-2.18 (m, 1H), 2.17-2.05(m, 4H), 1.94-1.77 (m, 2H), 1.51-1.39 (m, 4H), 1.25-1.08 (m, 5H), 0.86(t, J=7.3 Hz, 3H).

Preparation Example 1.36. (Dxd, Reference Example 5)

The reference compound (Dxd) was synthesized by referring to the methodprovided in Example 75 on page 183 of the specification of the patent“CN104755494A”.

The reference compound (Deruxtecan) was synthesized by referring to themethod provided in Example 58 on page 163 of the specification of thepatent “CN104755494A”. The reference ADC-1 (Trastuzumab-Deruxtecan) wassynthesized by referring to the method provided in Example 58 on page163 of the specification of the patent “CN104755494A” or by methodscommonly used in the art as disclosed.

Preparation Example 1.37

The following ligands can be prepared according to the conventionalmethods for antibodies. For example, vectors can be constructed,transfected into eukaryotic cells such as HEK293 cells (LifeTechnologies, Cat No. 11625019), and purified for expression.

Preparation Example 1.37a. The Sequences of Trastuzumab are Shown asFollows

Light chain (SEQ ID NO: 33)

DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC

Heavy chain (SEQ ID NO: 37)

EVQLVESGGGLVQPGGSLRLSCAASGFNKKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPG

Preparation Example 1.37b. The Sequences of Pertuzumab Antibody areShown as Follows

Light chain (SEQ ID NO: 34)

DIQMTQSPSSLSASVGDRVTITCKASQDVSIGVAWYQQKPGKAPKLLIYSASYRYTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYIYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC

Heavy chain (SEQ ID NO: 38)

EVQLVESGGGLVQPGGSLRLSCAASGFTFTDYTMDWVRQAPGKGLEWVADVNPNSGGSIYNQRFKGRFTLSVDRSKNTLYLQMNSLRAEDTAVYYCARNLGPSFYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQK SLSLSPG

Preparation Example 1.37c. The Sequences of hRS7 Antibody (Sacituzumab)are Shown as Follows

Light chain (SEQ ID NO: 35)

DIQLTQSPSSLSASVGDRVSITCKASQDVSIAVAWYQQKPGKAPKLLIYSASYRYTGVPDRFSGSGSGTDFTLTISSLQPEDFAVYYCQQHYITPLTFGAGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC

Heavy chain (SEQ ID NO: 39)

QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYTDDFKGRFAFSLDTSVSTAYLQISSLKADDTAVYFCARGGFGSSYWYFDVWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPGK

Preparation Example 1.37d. The Sequences of Zolbetuximab Antibody areShown as Follows

Light chain (SEQ ID NO: 36)

DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSYPFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS SPVTKSFNRGEC

Heavy chain (SEQ ID NO: 40)

QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWINWVKQRPGQGLEWIGNIYPSDSYTNYNQKFKDKATLTVDKSSSTAYMQLSSPTSEDSAVYYCTRSWRGNSFDYWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPGK

Preparation Example 1.38. ADC-II-1

The prepared tris(2-carboxyethyl)phosphine (5 mM, 0.233 mL, 1.163 μmol)aqueous solution was added to the antibody Trastuzumab in aqueous PBbuffer solution (0.04 M aqueous PB buffer solution at pH 7.0; 23 mg, 15mg/mL, 0.155 μmol) at 37° C., and the mixture was placed in a water bathshaker and shaken at 37° C. for 3 h, and then the reaction wasterminated;

L-II-1 (3.4 mg, 6.06 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution (2.0 mL), placed in a waterbath shaker and shaken at 25° C. for 3 h, and then the reaction wasterminated. The reaction solution was desalted and purified on aSephadex G25 gel column (eluent: 0.04 M aqueous PB buffer solution at pH7.0, containing 0.002 M EDTA) to give a solution of the exemplaryproduct ADC-II-1 in PB (3.08 mg/mL, 12 mg), which was stored at 4° C.

The mean n=7.42, as calculated by LC-MS.

Preparation Example 1.39. ADC-II-5

The prepared tris(2-carboxyethyl)phosphine (5 mM, 0.304 mL, 1.52 μmol)aqueous solution was added to the antibody Trastuzumab in aqueous PBbuffer solution (0.04 M aqueous PB buffer solution at pH 7.0; 30 mg, 15mg/mL, 0.203 μmol) at 37° C., and the mixture was placed in a water bathshaker and shaken at 37° C. for 3 h, and then the reaction wasterminated; the reaction solution was cooled to 25° C. in a water bathand diluted to 5.0 mg/mL, and the diluted solution (2.0 mL) was takenfor the next step.

L-II-2 (1.65 mg, 2.94 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution (2.0 mL), placed in a waterbath shaker and shaken at 25° C. for 3 h, and then the reaction wasterminated. The reaction solution was desalted and purified on aSephadex G25 gel column (eluent: 0.04 M aqueous PB buffer solution at pH7.0, containing 0.002 M EDTA) to give a solution of the exemplaryproduct ADC-II-5 in PB (6.37 mg/mL, 17 mg), which was frozen and storedat 4° C.

The mean n=7.58, as calculated by LC-MS.

Preparation Example 1.40. ADC-II-9

The prepared tris(2-carboxyethyl)phosphine (5 mM, 0.355 mL, 1.77 μmol)aqueous solution was added to the antibody trastuzumab in aqueous PBbuffer solution (0.04 M aqueous PB buffer solution at pH 7.0; 35 mg, 15mg/mL, 0.236 μmol) at 37° C., and the mixture was placed in a water bathshaker and shaken at 37° C. for 3 h, and then the reaction wasterminated; the reaction solution was cooled to 25° C. in a water bathand diluted to 5.0 mg/mL, and the diluted solution (2.0 mL) was takenfor the next step.

L-II-3 (4.51 mg, 2.84 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution (2.0 mL), placed in a waterbath shaker and shaken at 25° C. for 3 hours with shaking, and then thereaction was terminated. The reaction solution was desalted and purifiedon a Sephadex G25 gel column (eluent: 0.04 M aqueous PB buffer solutionat pH 7.0, containing 0.002 M EDTA) to give a solution of the exemplaryproduct ADC-II-9 in PB (7.00 mg/mL, 15 mg), which was frozen and storedat 4° C.

The mean n=8.05, as calculated by LC-MS.

Preparation Example 1.41. ADC-II-13

The prepared tris(2-carboxyethyl)phosphine (5 mM, 0.355 mL, 1.77 μmol)aqueous solution was added to the antibody Trastuzumab in aqueous PBbuffer solution (0.04 M aqueous PB buffer solution at pH 7.0; 35 mg, 15mg/mL, 0.236 μmol) at 37° C., and the mixture was placed in a water bathshaker and shaken at 37° C. for 3 h, and then the reaction wasterminated; the reaction solution was cooled to 25° C. in a water bathand diluted to 5.0 mg/mL, and the diluted solution (2.0 mL) was takenfor the next step.

L-II-4 (1.96 mg, 3.67 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution (2.0 mL), placed in a waterbath shaker and shaken at 25° C. for 3 h, and then the reaction wasterminated. The reaction solution was desalted and purified on aSephadex G25 gel column (eluent: 0.04 M aqueous PB buffer solution at pH7.0, containing 0.002 M EDTA) to give a solution of the exemplaryproduct ADC-III-13 in PB (6.94 mg/mL, 25 mg), which was frozen andstored at 4° C.

The mean n=7.89, as calculated by LC-MS.

Preparation Example 1.42. ADC-I-1

The prepared tris(2-carboxyethyl)phosphine (5 mM, 0.355 mL, 1.77 μmol)aqueous solution was added to the antibody Trastuzumab in aqueous PBbuffer solution (0.04 M aqueous PB buffer solution at pH 7.0; 35 mg, 15mg/mL, 0.236 μmol) at 37° C., and the mixture was placed in a water bathshaker and shaken at 37° C. for 3 h, and then the reaction wasterminated; the reaction solution was cooled to 25° C. in a water bathand diluted to 5.0 mg/mL, and the diluted solution (2.0 mL) was takenfor the next step.

L-I-1 (1.23 mg, 2.36 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution (2.0 mL), placed in a waterbath shaker and shaken at 25° C. for 3 h, and then the reaction wasterminated. The reaction solution was desalted and purified on aSephadex G25 gel column (eluent: 0.04 M aqueous PB buffer solution at pH7.0, containing 0.002 M EDTA) to give a solution of the exemplaryproduct ADC-I-1 in PB (9.33 mg/mL, 19 mg), which was frozen and storedat 4° C.

The mean n=7.98, as calculated by LC-MS.

Preparation Example 1.43. ADC-III-1

The prepared tris(2-carboxyethyl)phosphine (5 mM, 0.233 mL, 1.17 μmol)aqueous solution was added to the antibody Trastuzumab in PB buffer(0.04 M aqueous PB buffer solution at pH 7.0; 23 mg, 15 mg/mL, 0.155μmol) at 37° C., and the mixture was placed in a water bath shaker andshaken at 37° C. for 3 h, and then the reaction was terminated; thereaction solution was cooled to 25° C. in a water bath and diluted to5.0 mg/mL, and the diluted solution (2.0 mL) was taken for the nextstep.

L-III-2 (6.22 mg, 11.89 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution (2.0 mL), placed in a waterbath shaker and shaken at 25° C. for 3 h, and then the reaction wasterminated. The reaction solution was desalted and purified on aSephadex G25 gel column (eluent: 0.04 M aqueous PB buffer solution at pH7.0, containing 0.002 M EDTA) to give a solution of the exemplaryproduct ADC-III-1 in PB (1.25 mg/mL, 16 mg), which was frozen and storedat 4° C.

The mean n=8.34, as calculated by LC-MS.

Preparation Example 1.44. ADC-III-9

The prepared tris(2-carboxyethyl)phosphine (5 mM, 0.355 mL, 1.77 μmol)aqueous solution was added to the antibody Trastuzumab in aqueous PBbuffer solution (0.04 M aqueous PB buffer solution at pH 7.0; 35 mg, 15mg/mL, 0.236 μmol) at 37° C., and the mixture was placed in a water bathshaker and shaken at 37° C. for 3 h, and then the reaction wasterminated; the reaction solution was cooled to 25° C. in a water bathand diluted to 5.0 mg/mL, and the diluted solution (2.0 mL) was takenfor the next step.

L-III-20 (3.08 mg, 5.91 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution (2.0 mL), placed in a waterbath shaker and shaken at 25° C. for 3 h, and then the reaction wasterminated. The reaction solution was desalted and purified on aSephadex G25 gel column (eluent: 0.04 M aqueous PB buffer solution at pH7.0, containing 0.002 M EDTA) to give a solution of the exemplaryproduct ADC-III-9 in PB (6.61 mg/mL, 21 mg), which was frozen and storedat 4° C.

The mean n=8.05, as calculated by LC-MS.

Preparation Example 1.45. ADC-II-3

The prepared tris(2-carboxyethyl)phosphine (5 mM, 0.156 mL, 0.780 μmol)aqueous solution was added to the antibody Sacituzumab in aqueous PBbuffer solution (0.04 M aqueous PB buffer solution at pH 7.0; 35 mg, 11mg/mL, 0.236 μmol) at 25° C., and the mixture was placed in a water bathshaker and shaken at 25° C. for 3 h, and then the reaction wasterminated; the reaction solution was diluted to 5.0 mg/mL.

L-II-1 (2.60 mg, 2.36 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution, placed in a water bath shakerand shaken at 25° C. for 3 h, and then the reaction was terminated. Thereaction solution was desalted and purified on a Sephadex G25 gel column(eluent: 0.02 M aqueous histidine buffer solution at pH 5.5) to give asolution of the exemplary product ADC-II-3 in histidine (4.1 mg/mL, 22mg), which was refrigerated and stored at 4° C.

The mean n=3.89, as calculated by LC-MS.

Preparation Example 1.46. ADC-II-7

The prepared tris(2-carboxyethyl)phosphine (5 mM, 0.183 mL, 0.914 μmol)aqueous solution was added to the antibody Sacituzumab in PB buffer(0.04 M aqueous PB buffer solution at pH 7.0; 41 mg, 11 mg/mL, 0.277μmol) at 25° C., and the mixture was placed in a water bath shaker andshaken at 25° C. for 3 h, and then the reaction was terminated; thereaction solution was diluted to 5.0 mg/mL.

L-II-2 (3.05 mg, 2.77 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution, placed in a water bath shakerand shaken at 25° C. for 3 h, and then the reaction was terminated. Thereaction solution was desalted and purified on a Sephadex G25 gel column(eluent: 0.02 M aqueous histidine buffer solution at pH 5.5) to give asolution of the exemplary product ADC-II-7 in histidine (3.7 mg/mL, 29.5mg), which was refrigerated and stored at 4° C.

The mean n=4.12, as calculated by LC-MS.

Preparation Example 1.47. ADC-II-11-a

The prepared tris(2-carboxyethyl)phosphine (5 mM, 0.143 mL, 0.713 μmol)aqueous solution was added to the antibody Sacituzumab in PB buffer(0.04 M aqueous PB buffer solution at pH 7.0; 32 mg, 11 mg/mL, 0.216μmol) at 25° C., and the mixture was placed in a water bath shaker andshaken at 25° C. for 3 h, and then the reaction was terminated; thereaction solution was diluted to 5.0 mg/mL.

L-II-3 (2.32 mg, 2.16 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution, placed in a water bath shakerand shaken at 25° C. for 3 h, and then the reaction was terminated. Thereaction solution was desalted and purified on a Sephadex G25 gel column(eluent: 0.02 M aqueous histidine buffer solution at pH 5.5) to give asolution of the exemplary product ADC-II-11-a in histidine (2.2 mg/mL,15 mg), which was refrigerated and stored at 4° C.

The mean n=3.73, as calculated by LC-MS.

Preparation Example 1.48. ADC-II-15

The prepared tris(2-carboxyethyl)phosphine (5 mM, 0.165 mL, 0.825 μmol)aqueous solution was added to the antibody Sacituzumab in aqueous PBbuffer solution (0.04 M aqueous PB buffer solution at pH 7.0; 37 mg, 11mg/mL, 0.250 μmol) at 25° C., and the mixture was placed in a water bathshaker and shaken at 25° C. for 3 h, and then the reaction wasterminated; the reaction solution was diluted to 5.0 mg/mL.

L-II-4 (2.68 mg, 2.50 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution, placed in a water bath shakerand shaken at 25° C. for 3 h, and then the reaction was terminated. Thereaction solution was desalted and purified on a Sephadex G25 gel column(eluent: 0.02 M aqueous histidine buffer solution at pH 5.5) to give asolution of the exemplary product ADC-II-15 in histidine (4.0 mg/mL, 25mg), which was refrigerated and stored at 4° C.

The mean n=4.03, as calculated by LC-MS.

Preparation Example 1.49. ADC-III-3

The prepared tris(2-carboxyethyl)phosphine (5 mM, 0.156 mL, 0.780 μmol)aqueous solution was added to the antibody Sacituzumab in aqueous PBbuffer solution (0.04 M aqueous PB buffer solution at pH 7.0; 35 mg, 11mg/mL, 0.236 μmol) at 25° C., and the mixture was placed in a water bathshaker and shaken at 25° C. for 3 h, and then the reaction wasterminated; the reaction solution was diluted to 5.0 mg/mL.

L-III-2 (2.51 mg, 2.36 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution, placed in a water bath shakerand shaken at 25° C. for 3 h, and then the reaction was terminated. Thereaction solution was desalted and purified on a Sephadex G25 gel column(eluent: 0.02 M aqueous histidine buffer solution at pH 5.5) to give asolution of the exemplary product ADC-III-3 in histidine (2.7 mg/mL, 27mg), which was refrigerated and stored at 4° C.

The mean n=4.45, as calculated by LC-MS.

Preparation Example 1.50. ADC-III-11

The prepared tris(2-carboxyethyl)phosphine (5 mM, 0.174 mL, 0.870 μmol)aqueous solution was added to the antibody Sacituzumab in aqueous PBbuffer solution (0.04 M aqueous PB buffer solution at pH 7.0; 39 mg, 11mg/mL, 0.264 μmol) at 25° C., and the mixture was placed in a water bathshaker and shaken at 25° C. for 3 h, and then the reaction wasterminated;

L-III-20 (2.80 mg, 2.64 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution, placed in a water bath shakerand shaken at 25° C. for 3 h, and then the reaction was terminated. Thereaction solution was desalted and purified on a Sephadex G25 gel column(eluent: 0.02 M aqueous histidine buffer solution at pH 5.5) to give asolution of the exemplary product ADC-III-11 in histidine (7.89 mg/mL,31 mg), which was refrigerated and stored at 4° C.

The mean n=4.37, as calculated by LC-MS.

Preparation Example 1.50. ADC-II-11-b

The prepared tris(2-carboxyethyl)phosphine (5 mM, 0.089 mL, 0.446 μmol)aqueous solution was added to the antibody Sacituzumab in aqueous PBbuffer solution (0.04 M aqueous PB buffer solution at pH 7.0; 20 mg, 11mg/mL, 0.135 μmol) at 25° C., and the mixture was placed in a water bathshaker and shaken at 25° C. for 3 h, and then the reaction wasterminated; the reaction solution was diluted to 5.0 mg/mL.

L-II-3 (1.45 mg, 1.35 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution, placed in a water bath shakerand shaken at 25° C. for 3 h, and then the reaction was terminated. Thereaction solution was desalted and purified on a Sephadex G25 gel column(eluent: 0.02 M aqueous histidine buffer solution at pH 5.5) to give asolution of the exemplary product ADC-II-11-b in histidine (2.34 mg/mL,9.5 mg), which was refrigerated and stored at 4° C.

The mean n=8.02, as calculated by LC-MS.

Preparation Example 1.52. ADC-II-53

The prepared tris(2-carboxyethyl)phosphine (5 mM, 0.129 mL, 0.647 μmol)aqueous solution was added to the antibody Sacituzumab in aqueous PBbuffer solution (0.04 M aqueous PB buffer solution at pH 7.0; 29 mg, 11mg/mL, 0.196 μmol) at 25° C., and the mixture was placed in a water bathshaker and shaken at 25° C. for 3 h, and then the reaction wasterminated; the reaction solution was diluted to 5.0 mg/mL.

L-II-27 (1.82 mg, 1.96 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution, placed in a water bath shakerand shaken at 25° C. for 3 h, and then the reaction was terminated. Thereaction solution was desalted and purified on a Sephadex G25 gel column(eluent: 0.02 M aqueous histidine buffer solution at pH 5.5) to give asolution of the exemplary product ADC-II-53 in histidine (4.4 mg/mL,19.9 mg), which was refrigerated and stored at 4° C.

The mean n=4.46, as calculated by LC-MS.

Preparation Example 1.53. ADC-II-57

The prepared tris(2-carboxyethyl)phosphine (5 mM, 0.216 mL, 1.079 μmol)aqueous solution was added to the antibody Sacituzumab in aqueous PBbuffer solution (0.04 M aqueous PB buffer solution at pH 7.0; 48.4 mg,11 mg/mL, 0.327 μmol) at 25° C., and the mixture was placed in a waterbath shaker and shaken at 25° C. for 3 h, and then the reaction wasterminated; the reaction solution was diluted to 5.0 mg/mL.

L-II-28 (3.28 mg, 3.27 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution, placed in a water bath shakerand shaken at 25° C. for 3 h, and then the reaction was terminated. Thereaction solution was desalted and purified on a Sephadex G25 gel column(eluent: 0.02 M aqueous histidine buffer solution at pH 5.5) to give asolution of the exemplary product ADC-II-57 in histidine (4.15 mg/mL,33.4 mg), which was refrigerated and stored at 4° C.

The mean n=3.78, as calculated by LC-MS.

Preparation Example 1.54. ADC-III-28

The prepared tris(2-carboxyethyl)phosphine (5 mM, 0.251 mL, 1.255 μmol)aqueous solution was added to the antibody Sacituzumab in aqueous PBbuffer solution (0.04 M aqueous PB buffer solution at pH 7.0; 56.3 mg,11 mg/mL, 0.381 μmol) at 25° C., and the mixture was placed in a waterbath shaker and shaken at 25° C. for 3 h, and then the reaction wasterminated.

L-III-30 (4.05 mg, 3.81 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution, placed in a water bath shakerand shaken at 25° C. for 3 h, and then the reaction was terminated. Thereaction solution was desalted and purified on a Sephadex G25 gel column(eluent: 0.02 M aqueous histidine buffer solution at pH 5.5) to give asolution of the exemplary product ADC-III-28 in histidine (4.83 mg/mL,25.9 mg), which was refrigerated and stored at 4° C.

The mean n=3.67, as calculated by LC-MS.

Preparation Example 1.55. ADC-III-33

The prepared tris(2-carboxyethyl)phosphine (5 mM, 17.2 mL, 86.01 μmol)aqueous solution was added to the antibody Sacituzumab in aqueous PBbuffer solution (0.04 M aqueous PB buffer solution at pH 7.0; 3857 mg,11 mg/mL, 26.06 μmol) at 25° C., and the mixture was placed in a waterbath shaker and shaken at 25° C. for 3 h, and then the reaction wasterminated.

L-III-31 (4.05 mg, 3.81 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution, placed in a water bath shakerand shaken at 25° C. for 3 h, and then the reaction was terminated. Thereaction solution was desalted and purified on a Sephadex G25 gel column(eluent: 0.02 M aqueous histidine buffer solution at pH 5.5), andconcentrated in an Amicon ultrafiltration tube to give a solution of theexemplary product ADC-III-33 in histidine (23.71 mg/mL, 3433 mg), whichwas refrigerated and stored at 4° C.

The mean n=4.05, as calculated by LC-MS.

Preparation Example 1.56. Reference ADC-2 (Sacituzumab-Deruxtecan)

The prepared tris(2-carboxyethyl)phosphine (5 mM, 0.116 mL, 0.580 μmol)aqueous solution was added to the antibody Sacituzumab in aqueous PBbuffer solution (0.04 M aqueous PB buffer solution at pH 7.0; 26 mg, 11mg/mL, 0.176 μmol) at 25° C., and the mixture was placed in a water bathshaker and shaken at 25° C. for 3 h, and then the reaction wasterminated.

Deruxtecan (1.82 mg, 1.76 μmol) was dissolved in DMSO (0.10 mL), and thesolution was added to the above solution, placed in a water bath shakerand shaken at 25° C. for 3 h, and then the reaction was terminated. Thereaction solution was desalted and purified on a Sephadex G25 gel column(eluent: 0.02 M aqueous histidine buffer solution at pH 5.5) to give asolution of the reference ADC-2 in histidine (7.46 mg/mL, 23.6 mg),which was refrigerated and stored at 4° C.

The mean n=4.18, as calculated by LC-MS.

Example 2 Test Example 2.1. Test for Inhibition of In VitroProliferation of Tumor Cells by Compounds

Objective

To test the inhibitory activity of the pharmaceutical compounds on thein vitro proliferation of NCI-N87, JIMT-1 and MBA-MB-231 tumor cells.The cells were treated with the compounds at different concentrations invitro, and after 6 days of culture, the proliferation of cells wasdetected using the CTG (CellTiter-Glo® Luminescent Cell Viability Assay,Promega, Cat. No. G7558) reagent, and the in vitro activity of thecompounds was evaluated according to the IC₅₀ value.

Procedures

In the following, the test for the inhibition of the in vitroproliferation of NCI-N87 cells was taken as an example to illustrate themethod of the present application for testing the inhibitory activity ofthe compounds of the present application on the in vitro proliferationof tumor cells. The method was also applicable to, but not limited to,the test for the inhibitory activity on the in vitro proliferation ofother tumor cells.

1. Culturing of cells: NCI-N87 cells were cultured in RPMI-1640 mediumcontaining 10% FBS.

2. Preparation of cells: NCI-N87 cells in logarithmic phase were taken,washed with PBS once, and digested with 2-3 mL of trypsin for 2-3 min.After the cells were digested completely, 10-15 mL of cell culture wasadded to elute the digested cells. The eluate was centrifuged at 1000rpm for 5 min, and the supernatant was discarded. The resulting cellswere resuspended in 10-20 mL of cell culture to obtain a single cellsuspension.

3. Cell plating: the NCI-N87 single cell suspension was mixed well andadjusted to a viable cell density of 6×10⁴ cells/mL with cell culture.The cell suspension with the adjusted density was mixed well and addedto a 96-well cell culture plate at 50 μL/well. The culture plate wasincubated in an incubator for 18 h (37° C., 5% CO₂).

4. Preparation of compounds: the compounds were dissolved in DMSO toobtain stock solutions at an initial concentration of 10 mM.

There were 8 concentrations in total for the small molecule compounds:300 nM, 100 nM, 30 nM, 10 nM, 3 nM, 1 nM, 0.3 nM, and 0.1 nM.

5. Sample adding: the prepared samples to be detected at differentconcentrations were added to the culture plate, and two duplicate wellswere set for each sample. The culture plate was incubated in anincubator for 6 days (37° C., 5% CO₂).

6. Color developing: the 96-well cell culture plate was taken out, addedwith the CTG reagent at 50 μL/well, and incubated at room temperaturefor 10 min.

7. Plate reading: the 96-well cell culture plate was taken out, placedin a microplate reader, and measured for the chemiluminescence using themicroplate reader.

Data Analysis

Data were processed and analyzed using Microsoft Excel and GraphpadPrism 5.

TABLE 1 IC₅₀ values for the inhibition of the in vitro proliferation ofNCI-N87 and JIMT-1 cells by the small molecule fragments of the presentapplication. NCI-N87 JIMT-1 Compound No. IC₅₀ (nM) IC₅₀ (nM) P-II-118.47 8.47 P-II-2 22.1 17 P-II-3 9.558 4.996 P-II-4 8.032 6.637 P-I-197.29 23.7 P-I-1 61.56 21.81 P-III-2 108.4 108.1 P-III-20 5.888 4.351P-III-21 4.755 2.874

Conclusion: according to the results shown in Table 1, the smallmolecule fragments of the present application have significantinhibitory activity on the proliferation of NCI-N87 cells and JIMT-1cells. The compounds of the present application all have similarinhibitory activity on tumor proliferation.

Test Example 2.2. Test for Inhibition of In Vitro Proliferation of TumorCells by Compounds

Objective

To test the inhibitory activity of the pharmaceutical compounds on thein vitro proliferation of NCI-N87, JIMT-1 and MBA-MB-231 tumor cells.The cells were treated with compounds at different concentrations invitro, and after 6 days of culture, the proliferation of cells wasdetected using CTG (CellTiter-Glo® Luminescent Cell Viability Assay,Promega, Cat. No. G7558) reagents, and the in vitro activity of thecompounds was evaluated according to the IC₅₀ value.

1. Culturing of cells: NCI-N87/JIMT-1/MBA-MB-231 cells were cultured inRPMI-1640 medium containing 10% FBS.

2. Preparation of cells: NCI-N87/JIMT-1/MBA-MB-231 cells in logarithmicphase were taken, washed with PBS once, and digested with 2-3 mL oftrypsin for 2-3 min. After the cells were digested completely, 10-15 mLof cell culture was added to elute the digested cells. The eluate wascentrifuged at 1000 rpm for 5 min, and the supernatant was discarded.The resulting cells were resuspended in 10-20 mL of cell culture toobtain a single cell suspension.

3. Cell plating: the NCI-N87/JIMT-1/MBA-MB-231 single cell suspensionwas mixed well and adjusted to a viable cell density of 6×10⁴ cells/mLwith cell culture. The cell suspension with the adjusted density wasmixed well and added to a 96-well cell culture plate at 50 μL/well. Theculture plate was incubated in an incubator for 18 h (37° C., 5% CO₂).

4. Preparation of compounds: the compounds were dissolved in DMSO toobtain stock solutions at an initial concentration of 10 mM.

There were 8 concentrations in total for the small molecule compounds:300 nM, 100 nM, 30 nM, 10 nM, 3 nM, 1 nM, 0.3 nM, and 0.1 nM.

5. Sample adding: the prepared samples to be detected at differentconcentrations were added to the culture plate, and two duplicate wellswere set for each sample. The culture plate was incubated in anincubator for 6 days (37° C., 5% CO₂).

6. Color developing: the 96-well cell culture plate was taken out, addedwith the CTG reagent at 50 μL/well, and incubated at room temperaturefor 10 min.

7. Plate reading: the 96-well cell culture plate was taken out, placedin a microplate reader, and measured for the chemiluminescence using themicroplate reader.

Data analysis: data were processed and analyzed using Microsoft Exceland Graphpad Prism 5.

TABLE 2 IC₅₀ values for the inhibition of the in vitro proliferation oftumor cells by the small molecule fragments of the present application.NCI-N87 JIMT-1 MDA-MB-231 Compound No. IC₅₀ (nM) IC₅₀ (nM) IC₅₀ (nM)Reference — 23.1 — example 1 P-II-1 18.47 8.5 12.9 P-II-2 22.1 17.2 19.3P-II-3 9.558 5.0 9.4 P-II-4 8.032 6.6 7.6 P-II-22 — 3.2 — P-II-23 — —7.7 P-II-24 — 20.8 8.5

“-”: not detected

Conclusion: according to the results shown in Table 2, the smallmolecule fragments of the present application have significantinhibitory activity on the proliferation of NCI-N87, JIMT-1 andMDA-MB-231 cells. The compounds of the present application all havesimilar inhibitory activity on tumor proliferation.

Test Example 2.3. Test for Inhibition of In Vitro Proliferation of TumorCells by Compounds

Objective

To test the inhibitory activity of the pharmaceutical compounds on thein vitro proliferation of NCI-N87, JIMT-1 and MBA-MB-231 tumor cells.The cells were treated with compounds at different concentrations invitro, and after 6 days of culture, the proliferation of cells wasdetected using CTG (CellTiter-Glo® Luminescent Cell Viability Assay,Promega, Cat. No. G7558) reagents, and the in vitro activity of thecompounds was evaluated according to the IC₅₀ value.

1. Culturing of cells: NCI-N87/JIMT-1/MBA-MB-231 cells were cultured inRPMI-1640 medium containing 10% FBS.

2. Preparation of cells: NCI-N87/JIMT-1/MBA-MB-231 cells in logarithmicphase were taken, washed with PBS once, and digested with 2-3 mL oftrypsin for 2-3 min. After the cells were digested completely, 10-15 mLof cell culture was added to elute the digested cells. The eluate wascentrifuged at 1000 rpm for 5 min, and the supernatant was discarded.The resulting cells were resuspended in 10-20 mL of cell culture toobtain a single cell suspension.

3. Cell plating: the NCI-N87/JIMT-1/MBA-MB-231 single cell suspensionwas mixed well and adjusted to a viable cell density of 6×10⁴ cells/mLwith cell culture. The cell suspension with the adjusted density wasmixed well and added to a 96-well cell culture plate at 50 μL/well. Theculture plate was incubated in an incubator for 18 h (37° C., 5% CO₂).

4. Preparation of compounds: the compounds were dissolved in DMSO toobtain stock solutions at an initial concentration of 10 mM.

There were 8 concentrations in total for the small molecule compounds:300 nM, 100 nM, 30 nM, 10 nM, 3 nM, 1 nM, 0.3 nM, and 0.1 nM.

5. Sample adding: the prepared samples to be detected at differentconcentrations were added to the culture plate, and two duplicate wellswere set for each sample. The culture plate was incubated in anincubator for 6 days (37° C., 5% CO₂).

6. Color developing: the 96-well cell culture plate was taken out, addedwith the CTG reagent at 50 μL/well, and incubated at room temperaturefor 10 min.

7. Plate reading: the 96-well cell culture plate was taken out, placedin a microplate reader, and measured for the chemiluminescence using themicroplate reader.

Data analysis: data were processed and analyzed using Microsoft Exceland Graphpad Prism 5.

TABLE 3 IC₅₀ values for the inhibition of the in vitro proliferation ofNCI-N87, JIMT-1 and MDA-MB-231 cells by the small molecule fragments ofthe present application. NCI-N87 JIMT-1 MDA-MB-231 Compound No. IC50(nM) IC50 (nM) IC50 (nM) Reference — 36.2 — example 2 Reference — 81.2 —example 3 Reference — 32.3 — example 4 P-III-1 — — 28.9 P-III-2 108.4108.1 — P-III-9 — — 28.9 P-III-20 5.888 4.351 6.65 P-III-21 4.755 2.8742.88 P-III-22 — 12.5 — P-III-27 3.0 — — P-III-28 — — 1.2 P-III-29 — 16.78.6 P-III-30 — — 28.5

“-”: not detected

Conclusion: according to the results shown in Table 3, the smallmolecule fragments of the present application have significantinhibitory activity on the proliferation of NCI-N87, JIMT-1 andMDA-MB-231 cells. The compounds of the present application all havesimilar inhibitory activity on tumor proliferation.

Test Example 2.4. Test for Inhibition of In Vitro Proliferation of TumorCells by Compounds

Objective

To test the inhibitory activity of the pharmaceutical compounds on thein vitro proliferation of NCI-N87 and Colo205 tumor cells. The cellswere treated with compounds at different concentrations in vitro, andafter 6 days of culture, the proliferation of cells was detected usingCTG (CellTiter-Glo® Luminescent Cell Viability Assay, Promega, Cat. No.G7558) reagents, and the in vitro activity of the compounds wasevaluated according to the IC₅₀ value.

1. Culturing of cells: NCI-N87/Colo205 cells were cultured in RPMI-1640medium containing 10% FBS.

2. Preparation of cells: NCI-N87/Colo205 cells in logarithmic phase weretaken, washed with PBS once, and digested with 2-3 mL of trypsin for 2-3min. After the cells were digested completely, 10-15 mL of cell culturewas added to elute the digested cells. The eluate was centrifuged at1000 rpm for 5 min, and the supernatant was discarded. The resultingcells were resuspended in 10-20 mL of cell culture to obtain a singlecell suspension.

3. Cell plating: the NCI-N87/Colo205 single cell suspension was mixedwell and adjusted to a viable cell density of 6×10⁴ cells/mL with cellculture. The cell suspension with the adjusted density was mixed welland added to a 96-well cell culture plate at 50 μL/well. The cultureplate was incubated in an incubator for 18 h (37° C., 5% CO₂).

4. Preparation of compounds: the compounds were dissolved in DMSO toobtain stock solutions at an initial concentration of 10 mM.

There were 8 concentrations in total for the small molecule compounds:300 nM, 100 nM, 30 nM, 10 nM, 3 nM, 1 nM, 0.3 nM, and 0.1 nM.

5. Sample adding: the prepared samples to be detected at differentconcentrations were added to the culture plate, and two duplicate wellswere set for each sample. The culture plate was incubated in anincubator for 6 days (37° C., 5% CO₂).

6. Color developing: the 96-well cell culture plate was taken out, addedwith the CTG reagent at 50 μL/well, and incubated at room temperaturefor 10 min.

7. Plate reading: the 96-well cell culture plate was taken out, placedin a microplate reader, and measured for the chemiluminescence using themicroplate reader.

Data analysis: data were processed and analyzed using Microsoft Exceland Graphpad Prism 5.

TABLE 4 IC₅₀ values for the inhibition of the in vitro proliferation ofNCI-N87 and Colo205 cells by the small molecule fragments of the presentapplication. NCI-N87 Colo205 Compound No. IC50 (nM) IC50 (nM) P-III-205.9 20 P-III-30 3 9

Conclusion: according to the results shown in Table 4, the smallmolecule fragments of the present application have significantinhibitory activity on the proliferation of NCI-N87 cells and Co10205cells. The compounds of the present application all have similarinhibitory activity on tumor proliferation.

Test Example 2.5. Test for Inhibition of In Vitro Proliferation of TumorCells of HER2 Target by Antibody-Drug Conjugate

Objective

To test the inhibitory activity of the antibody drug conjugates againstthe HER2 target of the present application on the in vitro proliferationof NCI-N87, JIMT-1 and MBA-MB-231 tumor cells. The cells were treatedwith the ADCs at different concentrations in vitro, and after 6 days ofculture, the proliferation of cells was detected using the CTG reagent,and the in vitro activity of the compounds was evaluated according tothe IC₅₀ value.

Procedures

1. Culturing of cells: NCI-N87/JIMT-1 cells were cultured in RPMI-1640medium containing 10% FBS.

2. Preparation of cells: NCI-N87/JIMT-1 cells in logarithmic phase weretaken, washed with PBS once, and digested with 2-3 mL of trypsin for 2-3min. After the cells were digested completely, 10-15 mL of cell culturewas added to elute the digested cells. The eluate was centrifuged at1000 rpm for 5 min, and the supernatant was discarded. The resultingcells were resuspended in 10-20 mL of cell culture to obtain a singlecell suspension.

3. Cell plating: the NCI-N87/JIMT-1 single cell suspension was mixedwell and adjusted to a viable cell density of 6×10⁴ cells/mL with cellculture. The cell suspension with the adjusted density was mixed welland added to a 96-well cell culture plate at 50 μL/well. The cultureplate was incubated in an incubator for 18 h (37° C., 5% CO₂).

4. ADC concentration: the ADCs were diluted in a 5-fold gradientstarting from 150 nM to 0.000348 nM for a total of 9 concentrations.

5. Sample adding: the prepared samples to be detected at differentconcentrations were added to the culture plate, and two duplicate wellswere set for each sample. The culture plate was incubated in anincubator for 6 days (37° C., 5% CO₂).

6. Color developing: the 96-well cell culture plate was taken out, addedwith the CTG reagent at 50 μL/well, and incubated at room temperaturefor 10 min.

7. Plate reading: the 96-well cell culture plate was taken out, placedin a microplate reader, and measured for the chemiluminescence using themicroplate reader.

Data Analysis

Data were processed and analyzed using Microsoft Excel and GraphpadPrism 5.

TABLE 5 IC₅₀ values for the inhibition of the in vitro proliferation ofNCI-N87 and JIMT-1 cells by the ADCs of the present application. NCI-N87JIMT-1 ADC No. IC₅₀ (nM) IC₅₀ (nM) ADC-II-5 0.20 — ADC-II-9 0.08 39.91ADC-II-13 0.17 73.11 Reference ADC-1 0.28 90.55

Conclusion: according to the results shown in Table 5, the antibody-drugconjugates against the HER2 target of the present application havesignificant inhibitory activity on the proliferation of HER2-positivehigh expression cells NCI-N87, meanwhile, they have weak inhibitoryactivity on the proliferation of HER2-low expressing cells JIMT-1,therefore, they have good selectivity. The other ADCs of the presentapplication also have similar selective inhibitory activity.

Test Example 2.6. Test for Inhibition of In Vitro Proliferation of TumorCells of HER2 Target by Antibody-Drug Conjugate

Objective

To test the inhibitory activity of the antibody drug conjugates againstthe HER2 target of the present application on the in vitro proliferationof NCI-N87 and JIMT-1 tumor cells. The cells were treated with the ADCsat different concentrations in vitro, and after 6 days of culture, theproliferation of cells was detected using the CTG reagent, and the invitro activity of the compounds was evaluated according to the IC₅₀value.

Procedures

1. Culturing of cells: NCI-N87 or JIMT-1 cells were cultured inRPMI-1640 medium containing 10% FBS.

2. Preparation of cells: NCI-N87 or JIMT-1 cells in logarithmic phasewere taken, washed with PBS once, and digested with 2-3 mL of trypsinfor 2-3 min. After the cells were digested completely, 10-15 mL of cellculture was added to elute the digested cells. The eluate wascentrifuged at 1000 rpm for 5 min, and the supernatant was discarded.The resulting cells were resuspended in 10-20 mL of cell culture toobtain a single cell suspension.

3. Cell plating: the NCI-N87 or JIMT-1 single cell suspension was mixedwell and adjusted to a viable cell density of 6×10⁴ cells/mL with cellculture. The cell suspension with the adjusted density was mixed welland added to a 96-well cell culture plate at 50 μL/well. The cultureplate was incubated in an incubator for 18 h (37° C., 5% CO₂).

4. ADC concentration: The ADCs were diluted in a 5-fold gradientstarting from 150 nM to 0.000348 nM for a total of 9 concentrations.

5. Sample adding: the prepared samples to be detected at differentconcentrations were added to the culture plate, and two duplicate wellswere set for each sample. The culture plate was incubated in anincubator for 6 days (37° C., 5% CO₂).

6. Color developing: the 96-well cell culture plate was taken out, addedwith the CTG reagent at 50 μL/well, and incubated at room temperaturefor 10 min.

7. Plate reading: the 96-well cell culture plate was taken out, placedin a microplate reader, and measured for the chemiluminescence using themicroplate reader.

Data Analysis

Data were processed and analyzed using Microsoft Excel and GraphpadPrism 5.

TABLE 6 IC₅₀ values for the inhibition of the in vitro proliferation ofNCI-N87 and JIMT-1 cells by the ADCs of the present application. NCI-N87JIMT-1 ADC No. IC50 (nM) IC50 (nM) Reference ADC-1 0.28 90.55 ADC-II-10.14 — ADC-II-5 0.20 — ADC-II-9 0.095 28.1 ADC-II-13 0.17 73.11ADC-III-1 0.16 — ADC-III-9 0.13 76.7

Conclusion: according to the results shown in Table 6, the antibody-drugconjugates against the HER2 target of the present application havesignificant inhibitory activity on the proliferation of HER2-positivehigh expression cells NCI-N87, and they also have good inhibitoryactivity on the proliferation of HER2-low expressing cells JIMT-1. Theother ADCs of the present application also have similar inhibitoryactivity.

Test Example 2.7. Test for Inhibition of In Vitro Proliferation of TumorCells of Trop2 Target by Antibody-Drug Conjugate

Objective

To test the inhibitory activity of the antibody drug conjugates againstthe Trop2 target of the present application on the in vitroproliferation of Co10205 and SK-OV-3 tumor cells. The cells were treatedwith the ADCs at different concentrations in vitro, and after 6 days ofculture, the proliferation of cells was detected using the CTG reagent,and the in vitro activity of the compounds was evaluated according tothe IC₅₀ value.

Procedures

1. Culturing of cells: Colo205 or SK-OV-3 cells were cultured inRPMI-1640 medium containing 10% FBS.

2. Preparation of cells: Colo205 or SK-OV-3 cells in logarithmic phasewere taken, washed with PBS once, and digested with 2-3 mL of trypsinfor 2-3 min. After the cells were digested completely, 10-15 mL of cellculture was added to elute the digested cells. The eluate wascentrifuged at 1000 rpm for 5 min, and the supernatant was discarded.The resulting cells were resuspended in 10-20 mL of cell culture toobtain a single cell suspension.

3. Cell plating: the Colo205 or SK-OV-3 single cell suspension was mixedwell and adjusted to a viable cell density of 6×10⁴ cells/mL with cellculture. The cell suspension with the adjusted density was mixed welland added to a 96-well cell culture plate at 50 μL/well. The cultureplate was incubated in an incubator for 18 h (37° C., 5% CO₂).

4. ADC concentration: The ADCs were diluted in a 5-fold gradientstarting from 150 nM to 0.000348 nM for a total of 9 concentrations.

5. Sample adding: the prepared samples to be detected at differentconcentrations were added to the culture plate, and two duplicate wellswere set for each sample. The culture plate was incubated in anincubator for 6 days (37° C., 5% CO₂).

6. Color developing: the 96-well cell culture plate was taken out, addedwith the CTG reagent at 50 μL/well, and incubated at room temperaturefor 10 min.

7. Plate reading: the 96-well cell culture plate was taken out, placedin a microplate reader, and measured for the chemiluminescence using themicroplate reader.

Data Analysis

Data were processed and analyzed using Microsoft Excel and GraphpadPrism 5.

TABLE 7 IC₅₀ values for the inhibition of the in vitro proliferation ofColo205 and SK-OV-3 cells by the ADCs of the present application.Colo205 SK-OV-3 ADC No. IC50 (nM) IC50 (nM) Reference ADC-2 152.1 500.9ADC-II-11a 120.8 447.5 ADC-III-28 77.6 393.9

Conclusion: according to the results shown in Table 7, the antibody-drugconjugates against the Trop2 target of the present application havesignificant inhibitory activity on the proliferation of Trop2-positivehigh expression cells Co10205, meanwhile, they have weak inhibitoryactivity on the proliferation of HER2-low expressing cells JIMT-1,therefore, they have good selectivity. The other ADCs of the presentapplication also have similar selective inhibitory activity.

Test Example 2.8. Her2-ADC Plasma Stability Experiment

ADC-II-5, ADC-II-9 and ADC-II-13 at a final concentration of 1500 nMwere each added to sterile human plasma, and the resulting mixtures wereeach incubated in a cell incubator at 37° C., with the day of incubationrecorded as day 0, and then detected for free toxins at days 1, 7 and14.50 μL of each of the mixtures was taken for RP-HPLC analysis.

The detection results for free toxins show that ADC-II-5, ADC-II-9 andADC-II-13 are all quite stable in human plasma. The other ADCs of thepresent application also have similar stability.

Test Example 2.7. Her2-ADC Plasma Stability Experiment

Reference ADC-1, ADC-II-5, ADC-II-9 and ADC-II-13 at a finalconcentration of 1500 nM were each added to sterile human plasma, andthe resulting mixtures were each incubated in a cell incubator at 37°C., with the day of incubation recorded as day 0, and then detected forn value at 0 h, 3 h, 27 h and 99 h. 50 μL of each of the mixtures wastaken for LC-MS analysis.

TABLE 8 Test results of plasma stability for the exemplary ADCs in thepresent application n ADC No. T0 3 h 27 h 99 h Reference 7.56 6.47 5.633.55 ADC-1 ADC-II-5 7.58 7.57 7.18 4.69 ADC-II-9 8.05 7.89 7.48 6.40ADC-II-13 7.82 7.80 7.30 5.45

Conclusion: the results in Table 8 show that the conjugates formed bythe small molecule linkers of the present invention have significantlyless variation in n value in plasma than the reference ADC-1, showingsuperior plasma stability.

Test Example 2.9. Efficacy evaluation of NCI-N87 tumor-bearing mice

Objective

To evaluate the Her2-ADC antibodies of the present application usingBALB/c nude mice as test animals;

to evaluate the efficacy of ADC-II-5 and reference ADC-1 on NCI-N87xenograft tumor-bearing nude mice after the intraperitoneal injection.

Test compounds and materials

1. Test compounds

ADC-II-5: 0.5 mg/kg

ADC-II-5: 2 mg/kg

Reference ADC-1: 0.5 mg/kg

Reference ADC-1: 2 mg/kg

Blank control: PBS

2. Preparation method: all compounds were diluted with PBS.

3. Test animal

BALB/c Nude mice, purchased from Beijing Vital River.

Test Method

NCI-N87 cells were subcutaneously inoculated into the right flank of themice, and after tumors were grown for 8 days, animals were randomlygrouped, with 8 animals in each group for a total of 6 groups.

The compounds each were intraperitoneally injected once. The tumorvolume and weight were measured twice a week, and the data wererecorded.

Data were analyzed using Excel 2016 statistical software, wherein themean value was calculated as avg; the SD value was calculated as STDEV;the SEM value was calculated as STDEV/SQRT; and the P-value of thedifference between groups was calculated as TTEST.

The experimental results are shown, and it is observed from oneintraperitoneal injection to the end of the experiment that the ADCmolecules of the present application can significantly reduce the tumorvolume, and have superior tumor inhibition effect to the referenceADC-1. The other ADCs of the present application also have similar tumorinhibition effect in vivo.

Test Example 2.10. Efficacy Evaluation of NCI-N87 Tumor-Bearing Mice

Objective

To evaluate the conjugates of the present application using BALB/c nudemice as test animals;

to evaluate the efficacy of ADC-II-9, ADC-II-13, ADC-III-9 and referenceADC-1 on NCI-N₈₇ xenograft tumor-bearing nude mice after theintraperitoneal injection.

Test Compounds and Materials

1. Test compounds

ADC-II-9, ADC-II-13, ADC-III-9: 2 mg/kg

Reference ADC-1: 2 mg/kg

Blank control: PBS

2. Preparation method: all compounds were diluted with PBS.

3. Test animal

BALB/c Nude mice, purchased from Beijing Vital River.

Test Method

NCI-N87 cells were subcutaneously inoculated into the right flank of themice, and after tumors were grown for 8 days, animals were randomlygrouped, with 8 animals in each group for a total of 6 groups.

The compounds each were intraperitoneally injected once. The tumorvolume and weight were measured twice a week, and the data wererecorded.

Data were analyzed using Excel 2016 statistical software, wherein themean value was calculated as avg; the SD value was calculated as STDEV;the SEM value was calculated as STDEV/SQRT; and the P-value of thedifference between groups was calculated as TTEST.

TABLE 9 In vivo tumor inhibition effect of exemplary ADCs on the NCI-N87xenograft tumor model Tumor growth ADC No. inhibition % Reference ADC-154.14 ADC-II-9 134.29 ADC-II-13 58.75 ADC-III-9 93.79

The experimental results are shown in the table above and FIG. 1 , andit is observed from one intraperitoneal injection to the end of theexperiment that the ADC molecules of the present application cansignificantly reduce the tumor volume, and have superior tumorinhibition effect to the reference ADC-1. The other ADCs of the presentapplication also have similar tumor inhibition effect in vivo.

Test Example 2.11. Efficacy Evaluation of NCI-N87 Tumor-Bearing Mice

Objective

To evaluate the conjugates of the present application using BALB/c nudemice as test animals;

to evaluate the efficacy of ADC-II-9, ADC-III-1, ADC-III-9 and referenceADC-1 on NCI-N₈₇ xenograft tumor-bearing nude mice after theintraperitoneal injection.

Test Compounds and Materials

1. Test compounds

ADC-II-9, ADC-III-1, ADC-III-9: 2 mg/kg

Reference ADC-1: 2 mg/kg

Blank control: PBS

2. Preparation method: all compounds were diluted with PBS.

3. Test animal

BALB/c Nude mice, purchased from Beijing Vital River.

Test Method

NCI-N87 cells were subcutaneously inoculated into the right flank of themice, and after tumors were grown for 8 days, animals were randomlygrouped, with 8 animals in each group for a total of 6 groups.

The compounds each were intraperitoneally injected once. The tumorvolume and weight were measured twice a week, and the data wererecorded.

Data were analyzed using Excel 2016 statistical software, wherein themean value was calculated as avg; the SD value was calculated as STDEV;the SEM value was calculated as STDEV/SQRT; and the P-value of thedifference between groups was calculated as TTEST.

TABLE 10 In vivo tumor inhibition effect of exemplary ADCs on theNCI-N87 xenograft tumor model Tumor growth ADC No. inhibition %Reference ADC-1 54.14 ADC-II-9 68.86 ADC-III-1 80.39 ADC-III-9 93.05

The experimental results are shown in the table above and FIG. 2 , andit is observed from one intraperitoneal injection to the end of theexperiment that the ADC molecules of the present application cansignificantly reduce the tumor volume, and have superior tumorinhibition effect to the reference ADC-1. The other ADCs of the presentapplication also have similar tumor inhibition effect in vivo.

Test Example 2.12. Efficacy Evaluation of Colo205 Tumor-Bearing Mice

Objective

To evaluate the conjugates of the present application using BALB/c nudemice as test animals;

to evaluate the efficacy of ADC-III-28 and reference ADC-2 on Colo205xenograft tumor-bearing nude mice after the intraperitoneal injection.

Test Compounds and Materials

1. Test compounds

ADC-III-28: 10 mg/kg

Reference ADC-2: 10 mg/kg

Blank control: PBS

2. Preparation method: all compounds were diluted with PBS.

3. Test animal

BALB/c Nude mice, purchased from Beijing Vital River.

Test Method

Co10205 cells were subcutaneously inoculated into the right flank of themice, and after tumors were grown for 8 days, animals were randomlygrouped, with 8 animals in each group for a total of 6 groups.

The compounds each were intraperitoneally injected once. The tumorvolume and weight were measured twice a week, and the data wererecorded.

Data were analyzed using Excel 2016 statistical software, wherein themean value was calculated as avg; the SD value was calculated as STDEV;the SEM value was calculated as STDEV/SQRT; and the P-value of thedifference between groups was calculated as TTEST.

TABLE 11 In vivo tumor inhibition effect of exemplary ADCs on theColo205 xenograft tumor model Tumor growth ADC No. inhibition %Reference ADC-2 62.64 ADC-III-28 96.84

The experimental results are shown in the table above and FIG. 3 , andit is observed from one intraperitoneal injection to the end of theexperiment that the ADC molecules of the present application cansignificantly reduce the tumor volume, and have superior tumorinhibition effect to the reference ADC-2. The other ADCs of the presentapplication also have similar tumor inhibition effect in vivo.

Test Example 2.13. Efficacy Evaluation of SK-OV-3 Tumor-Bearing Mice

Objective

To evaluate the conjugates of the present application using BALB/c nudemice as test animals;

to evaluate the efficacy of ADC-III-28 and reference ADC-2 on SK-OV-3xenograft tumor-bearing nude mice after the intraperitoneal injection.

Test Compounds and Materials

1. Test compounds

ADC-III-28: 3 mg/kg, 10 mg/kg

Reference ADC-2: 3 mg/kg, 10 mg/kg

Blank control: PBS

2. Preparation method: all compounds were diluted with PBS.

3. Test animal

BALB/c Nude mice, purchased from Beijing Vital River.

Test Method

SK-OV-3 cells were subcutaneously inoculated into the right flank of themice, and after tumors were grown for 8 days, animals were randomlygrouped, with 8 animals in each group for a total of 6 groups.

The compounds each were intraperitoneally injected once. The tumorvolume and weight were measured twice a week, and the data wererecorded.

Data were analyzed using Excel 2016 statistical software, wherein themean value was calculated as avg; the SD value was calculated as STDEV;the SEM value was calculated as STDEV/SQRT; and the P-value of thedifference between groups was calculated as TTEST.

TABLE 12 In vivo tumor inhibition effect of exemplary ADCs on theSK-OV-3 xenograft tumor model Tumor growth ADC No. inhibition %Reference ADC-2 (3 mg/kg) 33.73% Reference ADC-2 (10 mg/kg) 55.01%ADC-III-28 (3 mg/kg) 58.74% ADC-III-28 (3 mg/kg) 81.18%

The experimental results are shown in the table above and FIG. 4 , andit is observed from one intraperitoneal injection to the end of theexperiment that the ADC molecules of the present application cansignificantly reduce the tumor volume, and have superior tumorinhibition effect to the reference ADC-2. The other ADCs of the presentapplication also have similar tumor inhibition effect in vivo.

Test Example 2.14. Efficacy Evaluation of NCI-N87 Tumor-Bearing Mice

Objective

To evaluate the conjugates of the present application using BALB/c nudemice as test animals;

to evaluate the efficacy of ADC-II-11-a, ADC-II-11-b, ADC-II-53,ADC-II-57 and ADC-III-28 on NCI-N87 xenograft tumor-bearing nude miceafter the intraperitoneal injection.

Test Compounds and Materials

1. Test compounds

ADC-II-11-a: 10 mg/kg

ADC-II-11-b: 10 mg/kg

ADC-II-53: 10 mg/kg

ADC-II-57: 10 mg/kg

ADC-III-28: 10 mg/kg

Blank control: PBS

2. Preparation method: all compounds were diluted with PBS.

3. Test animal

BALB/c Nude mice, purchased from Beijing Vital River.

Test Method

NCI-N87 cells were subcutaneously inoculated into the right flank of themice, and after tumors were grown for 8 days, animals were randomlygrouped, with 8 animals in each group for a total of 6 groups.

The compounds each were intraperitoneally injected once. The tumorvolume and weight were measured twice a week, and the data wererecorded.

Data were analyzed using Excel 2016 statistical software, wherein themean value was calculated as avg; the SD value was calculated as STDEV;the SEM value was calculated as STDEV/SQRT; and the P-value of thedifference between groups was calculated as TTEST.

The experimental results are shown in the table above and FIGS. 5, 6, 7,8, 9 and 10 , and it is observed from one intraperitoneal injection tothe end of the experiment that the ADC molecules of the presentapplication can significantly reduce the tumor volume. The other ADCs ofthe present application also have similar tumor inhibition effect invivo.

Test Example 2.15. Efficacy Evaluation of Colo205 Tumor-Bearing Mice

Objective

To evaluate the conjugates of the present application using BALB/c nudemice as test animals;

to evaluate the efficacy of ADC-III-28 and reference ADC-2 on Colo205xenograft tumor-bearing nude mice after the intraperitoneal injection.

Test Compounds and Materials

1. Test compounds

ADC-III-28: 10 mg/kg, one administration

Reference ADC-2: 10 mg/kg, two administrations

Blank control: PBS

2. Preparation method: all compounds were diluted with PBS.

3. Test animal

BALB/c Nude mice, purchased from Beijing Vital River.

Test Method

Colo205 cells were subcutaneously inoculated into the right flank of themice, and after tumors were grown for 8 days, animals were randomlygrouped, with 8 animals in each group for a total of 6 groups.

The compounds each were intraperitoneally injected once. The tumorvolume and weight were measured twice a week, and the data wererecorded.

Data were analyzed using Excel 2016 statistical software, wherein themean value was calculated as avg; the SD value was calculated as STDEV;the SEM value was calculated as STDEV/SQRT; and the P-value of thedifference between groups was calculated as TTEST.

The experimental results are shown in FIG. 10 , and it is observed fromone intraperitoneal injection to the end of the experiment thatADC-III-28 can significantly reduce the tumor volume as compared to thereference ADC-2 (two intraperitoneal injections), and has superior tumorinhibition effect to the reference ADC-2. The other ADCs of the presentapplication also have similar tumor inhibition effect in vivo.

Test Example 2.16. Efficacy Evaluation of JIMT-1 Tumor-Bearing Mice

Objective

To evaluate the conjugates of the present application using BALB/c nudemice as test animals;

to evaluate the efficacy of ADC-II-9 and reference ADC-1 on JIMT-1xenograft tumor-bearing nude mice after the intraperitoneal injection.

Test Compounds and Materials

1. Test compounds

ADC-II-9: 3 mg/kg, 10 mg/kg, one administration

Reference ADC-1: 3 mg/kg, 10 mg/kg, one administration

Blank control: PBS

2. Preparation method: all compounds were diluted with PBS.

3. Test animal

BALB/c Nude mice, purchased from Beijing Vital River.

Test Method

JIMT-1 cells were subcutaneously inoculated into the right flank of themice, and after tumors were grown for 8 days, animals were randomlygrouped, with 8 animals in each group for a total of 6 groups.

The compounds each were intraperitoneally injected once. The tumorvolume and weight were measured twice a week, and the data wererecorded.

Data were analyzed using Excel 2016 statistical software, wherein themean value was calculated as avg; the SD value was calculated as STDEV;the SEM value was calculated as STDEV/SQRT; and the P-value of thedifference between groups was calculated as TTEST.

The experimental results are shown in FIG. 11 , and it is observed fromone intraperitoneal injection to the end of the experiment that ADC-II-9can significantly reduce the tumor volume as compared to the referenceADC-1, and has superior tumor inhibition effect to the reference ADC-1.The other ADCs of the present application also have similar tumorinhibition effect in vivo.

Test Example 2.17. Efficacy Evaluation of Fadu Tumor-Bearing Mice

Objective

To evaluate the conjugates of the present application using BALB/c nudemice as test animals;

to evaluate the efficacy of ADC-II-11-a, ADC-II-11-b, ADC-II-53,ADC-II-57 and ADC-III-28 on Fadu xenograft tumor-bearing nude mice afterthe intraperitoneal injection.

Test Compounds and Materials

1. Test compounds

ADC-II-11-a: 10 mg/kg

ADC-II-11-b: 10 mg/kg

ADC-II-53: 10 mg/kg

ADC-II-57: 10 mg/kg

ADC-III-28: 10 mg/kg

Blank control: PBS

2. Preparation method: all compounds were diluted with PBS.

3. Test animal

BALB/c Nude mice, purchased from Beijing Vital River.

Test Method

Fadu cells were subcutaneously inoculated into the right flank of themice, and after tumors were grown for 8 days, animals were randomlygrouped, with 8 animals in each group for a total of 6 groups.

The compounds each were intraperitoneally injected once. The tumorvolume and weight were measured twice a week, and the data wererecorded.

Data were analyzed using Excel 2016 statistical software, wherein themean value was calculated as avg; the SD value was calculated as STDEV;the SEM value was calculated as STDEV/SQRT; and the P-value of thedifference between groups was calculated as TTEST.

The experimental results are shown in the table above and FIGS. 12, 13,14, 15 and 16 , and it is observed from one intraperitoneal injection tothe end of the experiment that the ADCs of the present invention all canreduce the tumor volume. The other ADCs of the present application alsohave similar tumor inhibition effect in vivo.

Example 3 Test Example 3.1. Bystander Effect

Objective

To investigate the bystander effect of the ADCs under the condition ofco-culture of HER2-positive tumor cells and HER2-negative tumor cells.

Test Method

Fluorescently labeled cells

Cells in the exponential phase were collected and viable cell werecounted using a cell counter.

The cell suspension was adjusted to a certain cell density with thecorresponding medium.

1 μL of each of Qtracker® components A and B was premixed in a 1.5 mL EPpipe to prepare the fluorescent labeling solutions. After incubation atroom temperature for 5 min, step 4) was performed immediately.

0.2 mL of fresh complete medium was added to the EP tube and vortexedfor 30 s.

1×10⁶ cells were added to the fluorescent labeling solution preparedabove.

The mixture was incubated at 37° C. for 60 min.

The cell labeling was examined under a fluorescent microscope.

The complete medium was added for washing twice. The resulting cellswere placed for later use.

Cell Inoculation

The 3 cell combinations as shown in Table 13 below were set up in a96-well cell culture plate:

TABLE 13 Cell combinations (+ representing the presence of a cell in thecell combination) Cell N87 HCC1187 number labeled Unlabeled labeledUnlabeled Group 1 + + − − Group 2 − + + − Group 3 − − + +

2) 90 μL of cell suspension was added to each well of a 96-well plate.

3) The 96-well plate was incubated in an incubator at 37° C. in 5% CO₂overnight.

3. Compound adding

1) A 10× drug dilution was prepared.

2) The diluted candidate molecules and positive drugs was added to eachwell, and 3 duplicate wells were set for each compound.

3) The 96-well plate was incubated in an incubator at 37° C. in 5% CO₂for 4 days.

4. Detection

1) Each set of images was observed and recorded under a fluorescencemicroscope.

2) A CTG solution which was pre-melted and equilibrated to roomtemperature was added to each well according to CTG operationinstructions, and the solution was mixed well using a microplate shaker,left to stand at room temperature for a period of time, and measured forfluorescence signal values using a plate reader.

3) Cell viability was expressed as mean fluorescence signal (treatmentgroup)/mean fluorescence signal (control group)×100%.

Under the experimental conditions, the ADC molecules of the presentapplication have a significant killing effect on HER2-positive cells,but have no significant inhibition effect on HER2-negative cells. Incells co-cultured using HER2-positive cells and HER2-negative cells, theADC molecules of the present application can significantly inhibit bothHER2-positive cells and HER2-negative cells, and show significantbystander effects. The other ADCs of the present application also havesimilar bystander effects.

Test Example 3.2. Study on Transporter Substrates

Objective

To investigate whether the compounds of the present invention is asubstrate for efflux transporters P-gp and BCRP in a Caco-2 cell model,thereby providing preclinical basis for analyzing the drug-druginteraction in pharmacokinetics.

Test method

Preparation of Monolayer Cells

Cell culture medium was added to each well of the Transwell. The HTSTranswell plate was incubated under conditions of 37° C. and 5% CO₂ for1 h prior to cell inoculation.

Caco-2 cells were diluted with culture medium and the cell suspensionwas dispensed into filter wells of the 96-well HTS Transwell plate.Cells were cultured under conditions of 37° C., 5% CO₂ and 95% relativehumidity for more than ten days. The cell culture medium was changedperiodically.

Monolayer cell resistance was measured using the Millicell EpithelialVolt-Ohm detection system. The resistance of each well was recorded.After all wells were measured, the Transwell plate was put back into theincubator.

TEER of each well was calculated according to the formula. The TEERvalue of each well should be greater than 230 ohm·cm².

Preparation for Transporter Test

The Caco-2 plate was taken out from the incubator. The monolayer cellswere washed with pre-heated HBSS twice. The plate was then incubated at37° C. for 30 min.

Stock solutions of the compounds were prepared in DMSO and diluted withHBSS to obtain working solutions. Digoxin was used as a referencesubstrate for P-gp and rosuvastatin was used as a reference substratefor BCRP. Propranolol was used as a high permeability marker.

The working solution (without inhibitor) was added to the Transwellinsert (apical compartment) to determine the rate of drug transport inthe apical to basolateral direction. The transfer buffer was added tothe wells in the receiver plate (basolateral compartment). The workingsolution (without inhibitor) was added to the receiver plate wells(basolateral compartment) to determine the rate of drug transport in thebasolateral to apical direction. The Transwell insert (apicalcompartment) was filled with the transport buffer. PSC833 was added tothe apical and basolateral compartments to determine the rate of drugtransport in the presence of P-gp inhibitors.

The sample was transferred from the working solution to the quenchingsolution, and a sample with a time of 0 was prepared. The Transwellplate was incubated under conditions of 37° C. and 5% CO₂ for 2 h.

At the end of the transport cycle, the samples were transferred from theapical and basolateral wells into a new 96-well plate. Cold acetonitrileor methanol containing the appropriate internal standard (IS) was addedto each well of the plate. The plate was vortexed for 10 min. Thesamples were centrifuged. The supernatant of an aliquot was mixed withan appropriate amount of ultrapure water (depending on the LC-MS/MSsignal response and peak shape) prior to the LC-MS/MS analysis.

The stock solution of lucifer yellow in DMSO was prepared and dilutedwith HBSS to determine the amount of lucifer yellow leakage after a2-hour transport period. The lucifer yellow solution was added to theapical compartment. HBSS was added to the basolateral compartment. Theplate was incubated at 37° C. for 30 min, and a certain amount of thesolution was directly taken from the apical and basolateral wells andtransferred to a new 96-well plate. The lucifer yellow fluorescence wasmeasured in a fluorescence plate reader with 485 nm excitation and 530nm emission (to monitor monolayer integrity).

Data Analysis

Data were calculated using Microsoft Excel. The percentage of thecompound remaining at each time point was estimated by determining thepeak area ratio from the extracted ion chromatogram.

For drug transport analysis in the Caco-2 cell model, the apparentpermeability coefficient (Papp) can be calculated according to thefollowing formula, in cm/s:Papp=(Creceiving side×Vreceiving side)/(Cinitiation×Tpenetrationtime×Smembrane area)

wherein the concentration C is in nM, the volume V is in μL, the time Tis in s, and the area S is in cm².

The recovery rate is calculated according to the following formula:

${{Recovery}{rate}} = \frac{\begin{matrix}{{{Creceiving}{side} \times {Vreceiving}{side}} + {{Ctadministrating}{side} \times}} \\{{{Vadministrating}{side}} + {{Ccell}{lysis}{buffer} \times V{cell}{lysis}{buffer}}}\end{matrix}}{{Cinitiation} \times {Vadministrating}{side}}$

The efflux ratio (ER) is calculated according to the following formula:

${{Efflux}{ratio}({ER})} = \frac{\left. {PappB}\rightarrow A \right.}{\left. {PappA}\rightarrow B \right.}$

The rate of change of the efflux ratio was calculated to evaluatewhether a compound is a substrate for P-gp or BCRP.

Test Results

Dxd is a transporter BCRP substrate but not a transporter P-gp substrateunder the test conditions as shown in Table 14. The test compoundsP-II-1, P-II-2, P-II-3, P-II-4 and P-I-1 are neither transporter P-gpsubstrates nor transporter BCRP substrates.

The results show that the compounds of the present application have alower risk of clinical drug-drug interactions than the reference drugDxd. The compounds of the present application all have similar drug-druginteractions.

TABLE 14 Study on transporter substrates of the compounds of the presentapplication P_(app) P_(app) _((A-B)) _((B-A)) Recovery Recovery (10⁻⁶,(10⁻⁶, Efflux rate (%) rate (%) BCRP Compound KO143 cm/s) cm/s) ratioAP-BL BL-AP substrate Propranolol − 17.30 18.92 1.09 62.16 82.36 NoRosuvastatin − 0.12 9.52 81.16 98.54 96.78 Yes + 0.45 3.15 6.96 95.8497.06 Dxd − 0.07 4.82 74.03 57.76 56.20 Yes + 0.21 4.42 21.23 76.5174.87 P-II-1 − 0.17 4.83 28.69 80.87 77.50 No + 0.20 5.52 27.54 76.9176.54 P-II-3 − 0.17 4.83 28.69 80.87 77.50 No + 0.20 5.52 27.54 76.9176.54 P-II-2 − 0.05 1.52 30.08 59.32 58.51 No + 0.07 1.70 22.74 62.4262.86 P-II-4 − 0.05 1.72 36.09 63.84 63.97 No + 0.06 1.65 25.47 65.0563.02 P-I-1 − 0.13 4.06 32.38 80.47 76.12 No + 0.21 4.94 23.49 81.4680.26

Test Example 3.3. Tissue Distribution Test in Tumor-Bearing Mice

Objective

After a single intravenous administration of ¹²⁵I isotopically labeledADC molecules of the present invention to mice bearing HER2-positivetumor cells, tissues/body fluids were collected at different time pointsfor y counting, and the tissue distribution profiles of the subjectswere researched according to the exposure level of the drug in differenttissues.

Test Method

The study on the tissue distribution of mice bearing HER2-positive tumorcells administered with the labeled subject ¹²⁵I-ADC was performed bythe ¹²⁵I isotope labeling method. In the test, the subject was firstlylabeled with [¹²⁵I] by an Iodogen method, the serum and tissue samplesof the ¹²⁵I-ADC were investigated by a TCA precipitation method, the[¹²⁵I]-labeled test sample was detected for biological activity, and ananimal test was conducted after the quality control of ¹²⁵I-ADC waspassed.

Mice (females) bearing HER2-positive tumor cells were administered with¹²⁵I-ADC, and various body fluids and tissue samples were collected atdifferent time points (6 animals at each time point) after theadministration. The total radioactivity and the radioactivityprecipitated after TCA precipitation were detected by a y counter toobtain the radioactive substance concentration in each tissue/bodyfluid, and the exposure of each tissue was calculated.

Test Results

Labeling results: the quality control after labeling was passed, and RCPof different batches of labeled samples placed in a solvent for acertain period of time meets the test requirements at 2-8° C.; thebiological activity after labeling meets the test requirements.

Results of tissue distribution test: the detection values of the totalthyroid radioactivity content at different time points after theadministration are all low, which does not affect the reliability of thetissue distribution test results.

After the administration to tumor-bearing mice, radioactivity isdistributed primarily in serum, secondarily in tumors, and partially orsparingly in other tissues and organs. The ADCs of the presentapplication all have similar tissue distribution profiles.

Test Example 3.4. Pharmacokinetic and Toxicity Studies of ADCs at aSingle Administration

Objective

To investigate the pharmacokinetic properties of the drug in monkeys andobserve the toxic manifestation of the animals after a singleintravenous drip of ADC in monkeys.

Test Method

Pharmacokinetics: after the single intravenous drip of ADC drugs atdifference doses in monkeys, blood samples were collected at a pluralityof continuous time points, and the concentration of the drugs in theblood was detected by a proper specific detection method.

Toxicity study: after the single intravenous drip of ADC drugs atdifference doses in monkeys, the tolerance of animals and drug-relatedtoxicity toxic manifestation were investigated in multiple aspects suchas clinical observation, body weight and food intake, hematology, bloodbiochemistry, urine and gross anatomy.

Test Results

After the single intravenous drip of ADCs in monkeys, the concentrationof free toxins is very low, and the pharmacokinetic properties of thetotal antibody and ADCs are similar, suggesting that ADCs are slowlyreleased in monkeys, have a stable conjugating mode, and can be used ina clinically planned administration frequency.

After the single intravenous drip of ADCs in monkeys, animals have goodtolerance and do not show serious or intolerable drug-related toxicity,suggesting that ADCs have controllable safety, and can be furtherstudied clinically. All ADCs of the present application have similarsafety.

The foregoing detailed description is provided by way of illustrationand example, and is not intended to limit the scope of the appendedclaims. Various modifications of the embodiments currently enumerated inthe present application will be apparent to those of ordinary skill inthe art and are intended to be within the scope of the appended claimsand their equivalents.

What is claimed is:
 1. A compound or a tautomer, a mesomer, a racemate,an enantiomer or a diastereoisomer thereof, or a mixture thereof, or apharmaceutically acceptable salt thereof, wherein the compound comprisesa structure shown as formula below:

wherein, R2 is —O—, and R2 is used for direct or indirect linking to aligand.
 2. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to claim 1,wherein the compound comprises a structure shown below:

wherein Ab is a ligand, the average connection number Na is an integeror a decimal from 1 to 10, and L is


3. The compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to claim 1,wherein the compound comprises a structure as shown below,

wherein, Ab is a ligand, and average connection n is an integer or adecimal from 1 to
 10. 4. The compound or the tautomer, the mesomer, theracemate, the enantiomer or the diastereoisomer thereof, or the mixturethereof, or the pharmaceutically acceptable salt thereof according toclaim 3, wherein the ligand Ab is an antibody.
 5. The compound or thetautomer, the mesomer, the racemate, the enantiomer or thediastereoisomer thereof, or the mixture thereof, or the pharmaceuticallyacceptable salt thereof according to claim 2 wherein the ligand Abtargets the following group: HER2, HER3, B7H3, TROP2, Claudin 18.2,CD30, CD33, CD70 and EGFR.
 6. The compound or the tautomer, the mesomer,the racemate, the enantiomer or the diastereoisomer thereof, or themixture thereof, or the pharmaceutically acceptable salt thereofaccording to claim 1, wherein the compound is selected from thefollowing structures:

wherein n is an integer or a decimal from 1 to
 10. 7. A pharmaceuticalcomposition, comprising the ligand-drug conjugate or the tautomer, themesomer, the racemate, the enantiomer or the diastereoisomer thereof, orthe mixture thereof, or the pharmaceutically acceptable salt thereofaccording to claim 1, and a pharmaceutically acceptable carrier.
 8. Amethod for treating tumor, comprising administering to a subject in needwith the compound or the tautomer, the mesomer, the racemate, theenantiomer or the diastereoisomer thereof, or the mixture thereof, orthe pharmaceutically acceptable salt thereof according to claim 1,and/or a pharmaceutical composition that may comprise the same.